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Sample records for agrobacterium-mediated transformation method

  1. ipt Gene transformation in petunia by an Agrobacterium mediated method.

    PubMed

    Bai, L J; Ye, C J; Lu, J Y; Yang, D E; Xue, H; Pan, Y; Cao, P X; Wang, B; Liu, M

    2009-01-01

    To prevent leaf senescence of petunia, the cytokinin biosynthetic gene isopentenyl transferase (ipt) was placed under the control of 35S promoter and introduced into petunia. PCR analysis showed an expected 0.5 Kb fragment of ipt gene in transgenic petunia. RT-PCR analysis indicated the expression of ipt gene in the transgenic lines. Leaves from transgenic plants remained green and healthy in normal culture condition, while the non-transformed plants turned to yellow. Transgenic plants showed a reduction in height and smaller leaf sizes. In transgenic lines, the internodes were shorter, and the roots grew slower than the non-transformed plants.

  2. Barley Transformation Using Agrobacterium-Mediated Techniques

    NASA Astrophysics Data System (ADS)

    Harwood, Wendy A.; Bartlett, Joanne G.; Alves, Silvia C.; Perry, Matthew; Smedley, Mark A.; Leyland, Nicola; Snape, John W.

    Methods for the transformation of barley using Agrobacterium-mediated techniques have been available for the past 10 years. Agrobacterium offers a number of advantages over biolistic-mediated techniques in terms of efficiency and the quality of the transformed plants produced. This chapter describes a simple system for the transformation of barley based on the infection of immature embryos with Agrobacterium tumefaciens followed by the selection of transgenic tissue on media containing the antibiotic hygromycin. The method can lead to the production of large numbers of fertile, independent transgenic lines. It is therefore ideal for studies of gene function in a cereal crop system.

  3. A rapid, highly efficient and economical method of Agrobacterium-mediated in planta transient transformation in living onion epidermis.

    PubMed

    Xu, Kedong; Huang, Xiaohui; Wu, Manman; Wang, Yan; Chang, Yunxia; Liu, Kun; Zhang, Ju; Zhang, Yi; Zhang, Fuli; Yi, Liming; Li, Tingting; Wang, Ruiyue; Tan, Guangxuan; Li, Chengwei

    2014-01-01

    Transient transformation is simpler, more efficient and economical in analyzing protein subcellular localization than stable transformation. Fluorescent fusion proteins were often used in transient transformation to follow the in vivo behavior of proteins. Onion epidermis, which has large, living and transparent cells in a monolayer, is suitable to visualize fluorescent fusion proteins. The often used transient transformation methods included particle bombardment, protoplast transfection and Agrobacterium-mediated transformation. Particle bombardment in onion epidermis was successfully established, however, it was expensive, biolistic equipment dependent and with low transformation efficiency. We developed a highly efficient in planta transient transformation method in onion epidermis by using a special agroinfiltration method, which could be fulfilled within 5 days from the pretreatment of onion bulb to the best time-point for analyzing gene expression. The transformation conditions were optimized to achieve 43.87% transformation efficiency in living onion epidermis. The developed method has advantages in cost, time-consuming, equipment dependency and transformation efficiency in contrast with those methods of particle bombardment in onion epidermal cells, protoplast transfection and Agrobacterium-mediated transient transformation in leaf epidermal cells of other plants. It will facilitate the analysis of protein subcellular localization on a large scale.

  4. A Rapid, Highly Efficient and Economical Method of Agrobacterium-Mediated In planta Transient Transformation in Living Onion Epidermis

    PubMed Central

    Xu, Kedong; Huang, Xiaohui; Wu, Manman; Wang, Yan; Chang, Yunxia; Liu, Kun; Zhang, Ju; Zhang, Yi; Zhang, Fuli; Yi, Liming; Li, Tingting; Wang, Ruiyue; Tan, Guangxuan; Li, Chengwei

    2014-01-01

    Transient transformation is simpler, more efficient and economical in analyzing protein subcellular localization than stable transformation. Fluorescent fusion proteins were often used in transient transformation to follow the in vivo behavior of proteins. Onion epidermis, which has large, living and transparent cells in a monolayer, is suitable to visualize fluorescent fusion proteins. The often used transient transformation methods included particle bombardment, protoplast transfection and Agrobacterium-mediated transformation. Particle bombardment in onion epidermis was successfully established, however, it was expensive, biolistic equipment dependent and with low transformation efficiency. We developed a highly efficient in planta transient transformation method in onion epidermis by using a special agroinfiltration method, which could be fulfilled within 5 days from the pretreatment of onion bulb to the best time-point for analyzing gene expression. The transformation conditions were optimized to achieve 43.87% transformation efficiency in living onion epidermis. The developed method has advantages in cost, time-consuming, equipment dependency and transformation efficiency in contrast with those methods of particle bombardment in onion epidermal cells, protoplast transfection and Agrobacterium-mediated transient transformation in leaf epidermal cells of other plants. It will facilitate the analysis of protein subcellular localization on a large scale. PMID:24416168

  5. Agrobacterium-mediated sorghum transformation.

    PubMed

    Zhao, Z Y; Cai, T; Tagliani, L; Miller, M; Wang, N; Pang, H; Rudert, M; Schroeder, S; Hondred, D; Seltzer, J; Pierce, D

    2000-12-01

    Agrobacterium tumefaciens was used to genetically transform sorghum. Immature embryos of a public (P898012) and a commercial line (PHI391) of sorghum were used as the target explants. The Agrobacterium strain used was LBA4404 carrying a 'Super-binary' vector with a bar gene as a selectable marker for herbicide resistance in the plant cells. A series of parameter tests was used to establish a baseline for conditions to be used in stable transformation experiments. A number of different transformation conditions were tested and a total of 131 stably transformed events were produced from 6175 embryos in these two sorghum lines. Statistical analysis showed that the source of the embryos had a very significant impact on transformation efficiency, with field-grown embryos producing a higher transformation frequency than greenhouse-grown embryos. Southern blot analysis of DNA from leaf tissues of T0 plants confirmed the integration of the T-DNA into the sorghum genome. Mendelian segregation in the T1 generation was confirmed by herbicide resistance screening. This is the first report of successful use of Agrobacterium for production of stably transformed sorghum plants. The Agrobacterium method we used yields a higher frequency of stable transformation that other methods reported previously.

  6. An efficient method for Agrobacterium-mediated genetic transformation and plant regeneration in cumin (Cuminum cyminum L.).

    PubMed

    Pandey, Sonika; Mishra, Avinash; Patel, Manish Kumar; Jha, Bhavanath

    2013-09-01

    Cumin is an annual herbaceous medicinally important plant having diverse applications. An efficient and reproducible method of Agrobacterium-mediated genetic transformation was herein established for the first time. A direct regeneration method without callus induction was optimised using embryos as explant material in Gamborg's B5 medium supplemented with 0.5-μM 6-benzyladenine and 2.0-μM α-naphthalene acetic acid. About 1,020 embryos (a mean of 255 embryos per batch) were used for the optimisation of transformation conditions. These conditions were an Agrobacterium cell suspension of 0.6 OD600, a co-cultivation time of 72 h, 300-μM acetosyringone and wounding of explants using a razor blade. Pre-cultured elongated embryos were treated using optimised conditions. About 720 embryos (a mean of 180 embryos per batch) were used for transformation and 95 % embryos showed transient β-glucuronidase expression after co-cultivation. Putative transformed embryos were cultured on B5 medium for shoot proliferation and 21 regenerated plants were obtained after selection and allowed to root. T0 plantlets showed β-glucuronidase expression and gene integration was confirmed via PCR amplification of 0.96 and 1.28 kb fragments of the hygromycin-phosphotransferase II and β-glucuronidase genes, respectively. In this study, a transformation efficiency of 1.5 % was demonstrated and a total of 11 transgenic plants were obtained at the hardening stage, however, only four plants acclimatised during hardening. Gene copy number was analysed by Southern blot analysis of hardened plants and single-copy gene integration was observed. This is the first successful attempt of Agrobacterium-mediated genetic transformation of cumin.

  7. Development of an Agrobacterium-Mediated Transformation Method and Evaluation of Two Exogenous Constitutive Promoters in Oleaginous Yeast Lipomyces starkeyi.

    PubMed

    Lin, Xinping; Liu, Sasa; Bao, Ruiqi; Gao, Ning; Zhang, Sufang; Zhu, Rongqian; Zhao, Zongbao Kent

    2017-04-06

    Oleaginous yeast Lipomyces starkeyi, a promising strain of great biotechnical importance, is able to accumulate over 60% of its cell biomass as triacylglycerols (TAGs). It is promising to directly produce the derivatives of TAGs, such as long-chain fatty acid methyl esters and alkanes, in L. starkeyi. However, techniques for genetic modification of this oleaginous yeast are lacking, thus, further research is needed to develop genetic tools and functional elements. Here, we used two exogenous promoters (pGPD and pPGK) from oleaginous yeast Rhodosporidium toruloides to establish a simpler Agrobacterium-mediated transformation (AMT) method for L. starkeyi. Hygromycin-resistant transformants were obtained on antibiotic-contained plate. Mitotic stability test, genotype verification by PCR, and protein expression confirmation all demonstrated the success of this method. Furthermore, the strength of these two promoters was evaluated at the phenotypic level on a hygromycin-gradient plate and at the transcriptional level by real-time quantitative PCR. The PGK promoter strength was 2.2-fold as that of GPD promoter to initiate the expression of the hygromycin-resistance gene. This study provided an easy and efficient genetic manipulation method and elements of the oleaginous yeast L. starkeyi for constructing superior strains to produce advanced biofuels.

  8. Agrobacterium-mediated transformation of Fusarium proliferatum.

    PubMed

    Bernardi-Wenzel, J; Quecine, M C; Azevedo, J L; Pamphile, J A

    2016-06-03

    Fusarium proliferatum is an important pathogen that is associated with plant diseases and primarily affects aerial plant parts by producing different mycotoxins, which are toxic to humans and animals. Within the last decade, this fungus has also been described as one of the causes of red root rot or sudden death syndrome in soybean, which causes extensive damage to this crop. This study describes the Agrobacterium tumefaciens-mediated transformation of F. proliferatum as a tool for the disruption of pathogenicity genes. The genetic transformation was performed using two binary vectors (pCAMDsRed and pFAT-GFP) containing the hph (hygromycin B resistance) gene as a selection marker and red and green fluorescence, respectively. The presence of acetosyringone and the use of filter paper or nitrocellulose membrane were evaluated for their effect on the transformation efficiency. A mean processing rate of 94% was obtained with 96 h of co-cultivation only in the presence of acetosyringone and the use of filter paper or nitrocellulose membrane did not affect the transformation process. Hygromycin B resistance and the presence of the hph gene were confirmed by PCR, and fluorescence due to the expression of GFP and DsRed protein was monitored in the transformants. A high rate of mitotic stability (95%) was observed. The efficiency of Agrobacterium-mediated transformation of F. proliferatum allows the technique to be used for random insertional mutagenesis studies and to analyze fungal genes involved in the infection process.

  9. Agrobacterium-mediated transformation of maize (Zea mays) immature embryos.

    PubMed

    Lee, Hyeyoung; Zhang, Zhanyuan J

    2014-01-01

    Agrobacterium tumefaciens-mediated transformation is one of the most efficient and simple gene delivery systems for genetic improvement and biology studies in maize. This system has become more widely used by both public and private laboratories. However, transformation efficiencies vary greatly from laboratory to laboratory for the same genotype. Here, we illustrate our advanced Agrobacterium-mediated transformation method in Hi-II maize using simple binary vectors. The protocol utilizes immature embryos as starting explants and the bar gene as a selectable marker coupled with bialaphos as a selective agent. The protocol offers efficient transformation results with high reproducibility, provided that some experimental conditions are well controlled. This transformation method, with minor modifications, can be also employed to transform certain maize inbreds.

  10. Optimization of Agrobacterium-Mediated Transformation in Soybean

    PubMed Central

    Li, Shuxuan; Cong, Yahui; Liu, Yaping; Wang, Tingting; Shuai, Qin; Chen, Nana; Gai, Junyi; Li, Yan

    2017-01-01

    High transformation efficiency is a prerequisite for study of gene function and molecular breeding. Agrobacterium tumefaciens-mediated transformation is a preferred method in many plants. However, the transformation efficiency in soybean is still low. The objective of this study is to optimize Agrobacterium-mediated transformation in soybean by improving the infection efficiency of Agrobacterium and regeneration efficiency of explants. Firstly, four factors affecting Agrobacterium infection efficiency were investigated by estimation of the rate of GUS transient expression in soybean cotyledonary explants, including Agrobacterium concentrations, soybean explants, Agrobacterium suspension medium, and co-cultivation time. The results showed that an infection efficiency of over 96% was achieved by collecting the Agrobacterium at a concentration of OD650 = 0.6, then using an Agrobacterium suspension medium containing 154.2 mg/L dithiothreitol to infect the half-seed cotyledonary explants (from mature seeds imbibed for 1 day), and co-cultured them for 5 days. The Agrobacterium infection efficiencies for soybean varieties Jack Purple and Tianlong 1 were higher than the other six varieties. Secondly, the rates of shoot elongation were compared among six different concentration combinations of gibberellic acid (GA3) and indole-3-acetic acid (IAA). The shoot elongation rate of 34 and 26% was achieved when using the combination of 1.0 mg/L GA3 and 0.1 mg/L IAA for Jack Purple and Tianlong 1, respectively. This rate was higher than the other five concentration combinations of GA3 and IAA, with an 18 and 11% increase over the original laboratory protocol (a combination of 0.5 mg/L GA3 and 0.1 mg/L IAA), respectively. The transformation efficiency was 7 and 10% for Jack Purple and Tianlong 1 at this optimized hormone concentration combination, respectively, which was 2 and 6% higher than the original protocol, respectively. Finally, GUS histochemical staining, PCR, herbicide

  11. An efficient method of agrobacterium-mediated genetic transformation and regeneration in local Indian cultivar of groundnut (Arachis hypogaea) using grafting.

    PubMed

    Tiwari, Vivekanand; Chaturvedi, Amit Kumar; Mishra, Avinash; Jha, Bhavanath

    2015-01-01

    Groundnut (Arachis hypogaea L.) is an industrial crop used as a source of edible oil and nutrients. In this study, an efficient method of regeneration and Agrobacterium-mediated genetic transformation is reported for a local cultivar GG-20 using de-embryonated cotyledon explant. A high regeneration 52.69 ± 2.32 % was achieved by this method with 66.6 μM 6-benzylaminopurine (BAP), while the highest number of shoot buds per explant, 17.67 ± 3.51, was found with 20 μM BAP and 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D). The bacterial culture OD, acetosyringone and L-cysteine concentration were optimized as 1.8, 200 μM and 50 mg L(-1), respectively, in co-cultivation media. It was observed that the addition of 2,4-D in co-cultivation media induced accumulation of endogenous indole-3-acetic acid (IAA). The optimized protocol exhibited 85 % transformation efficiency followed by 14.65 ± 1.06 % regeneration, of which 3.82 ± 0.6 % explants were survived on hygromycin after selection. Finally, 14.58 ± 2.95 % shoots (regenerated on survived explants) were rooted on rooting media (RM3). In grafting method, regenerated shoots (after hygromycin selection) were grafted on the non-transformed stocks with 100 % survival and new leaves emerged in 3 weeks. The putative transgenic plants were then confirmed by PCR, Southern hybridization, reverse transcriptase PCR (RT-PCR) and β-glucuronidase (GUS) histochemical assay. The reported method is efficient and rapid and can also be applied to other crops which are recalcitrant and difficult in rooting.

  12. Lox-dependent gene expression in transgenic plants obtained via Agrobacterium-mediated transformation.

    PubMed

    Shcherbak, N; Kishchenko, O; Sakhno, L; Komarnytsky, I; Kuchuk, M

    2013-01-01

    Lox sites of the Cre/lox recombination system from bacteriophage P1 were analyzed for their ability to affect on transgene expression when inserted upstream from a gene coding sequence adjacent to the right border (RB) of T-DNA. Wild and mutated types of lox sites were tested for their effect upon bar gene expression in plants obtained via Agrobacterium-mediated and biolistic transformation methods. Lox-mediated expression of bar gene, recognized by resistance of transgenic plants to PPT, occurred only in plants obtained via Agrobacterium-mediated transformation. RT-PCR analysis confirms that PPT-resistant phenotype of transgenic plants obtained via Agrobacterium-mediated transformation was caused by activation of bar gene. The plasmid with promoterless gus gene together with the lox site adjacent to the RB was constructed and transferred to Nicotiana tabacum as well. Transgenic plants exhibited GUS activity and expression of gus gene was detected in plant leaves. Expression of bar gene from the vectors containing lox site near RB allowed recovery of numerous PPT-resistant transformants of such important crops as Beta vulgaris, Brassica napus, Lactuca sativa and Solanum tuberosum. Our results demonstrate that the lox site sequence adjacent to the RB can be used to control bar gene expression in transgenic plants.

  13. Plant Cell Division Analyzed by Transient Agrobacterium-Mediated Transformation of Tobacco BY-2 Cells.

    PubMed

    Buschmann, Henrik

    2016-01-01

    The continuing analysis of plant cell division will require additional protein localization studies. This is greatly aided by GFP-technology, but plant transformation and the maintenance of transgenic lines can present a significant technical bottleneck. In this chapter I describe a method for the Agrobacterium-mediated genetic transformation of tobacco BY-2 cells. The method allows for the microscopic analysis of fluorescence-tagged proteins in dividing cells in within 2 days after starting a coculture. This transient transformation procedure requires only standard laboratory equipment. It is hoped that this rapid method would aid researchers conducting live-cell localization studies in plant mitosis and cytokinesis.

  14. Agrobacterium-mediated transformation of bottle gourd (Lagenaria siceraria Standl.).

    PubMed

    Han, J-S; Kim, C K; Park, S H; Hirschi, K D; Mok, I- G

    2005-03-01

    We describe a procedure for producing transgenic bottle gourd plants by inoculating cotyledon explants with Agrobacterium tumefaciens strain AGL1 that carries the binary vector pCAMBIA3301 containing a glufosinate ammonium-resistance (bar) gene and the beta-D-glucuronidase (GUS) reporter gene. The most effective bacterial infection was observed when cotyledon explants of 4-day-old seedlings were co-cultivated with Agrobacterium for 6-8 days on co-cultivation medium supplemented with 0.1-0.001 mg/l L-alpha-(2-aminoethoxyvinyl) glycine (AVG). The putatively transformed shoots directly emerged at the proximal end of cotyledon explants after 2-3 weeks of culturing on selection medium containing 2 mg/l DL-phosphinothricin. These shoots were rooted after 3 weeks of culturing on half-strength MS medium containing 0.1 mg/l indole acetic acid and 1 mg/l DL-phosphinothricin. Transgenic plants were obtained at frequencies of 1.9%. Stable integration and transmission of the transgenes in T1 generation plants were confirmed by a histochemical GUS assay, polymerase chain reaction and Southern blot analyses. Genetic segregation analysis of T1 progenies showed that transgenes were inherited in a Mendelian fashion. To our knowledge, this study is the first to show Agrobacterium-mediated transformation in bottle gourd.

  15. Genomic regions responsible for amenability to Agrobacterium-mediated transformation in barley

    PubMed Central

    Hisano, Hiroshi; Sato, Kazuhiro

    2016-01-01

    Different plant cultivars of the same genus and species can exhibit vastly different genetic transformation efficiencies. However, the genetic factors underlying these differences in transformation rate remain largely unknown. In barley, ‘Golden Promise’ is one of a few cultivars reliable for Agrobacterium-mediated transformation. By contrast, cultivar ‘Haruna Nijo’ is recalcitrant to genetic transformation. We identified genomic regions of barley important for successful transformation with Agrobacterium, utilizing the ‘Haruna Nijo’ × ‘Golden Promise’ F2 generation and genotyping by 124 genome-wide SNP markers. We observed significant segregation distortions of these markers from the expected 1:2:1 ratio toward the ‘Golden Promise’-type in regions of chromosomes 2H and 3H, indicating that the alleles of ‘Golden Promise’ in these regions might contribute to transformation efficiency. The same regions, which we termed Transformation Amenability (TFA) regions, were also conserved in transgenic F2 plants generated from a ‘Morex’ × ‘Golden Promise’ cross. The genomic regions identified herein likely include necessary factors for Agrobacterium-mediated transformation in barley. The potential to introduce these loci into any haplotype of barley opens the door to increasing the efficiency of transformation for target alleles into any haplotype of barley by the TFA-based methods proposed in this report. PMID:27874056

  16. Improved methods in Agrobacterium-mediated transformation of almond using positive (mannose/pmi) or negative (kanamycin resistance) selection-based protocols.

    PubMed

    Ramesh, Sunita A; Kaiser, Brent N; Franks, Tricia; Collins, Graham; Sedgley, Margaret

    2006-08-01

    A protocol for Agrobacterium-mediated transformation with either kanamycin or mannose selection was developed for leaf explants of the cultivar Prunus dulcis cv. Ne Plus Ultra. Regenerating shoots were selected on medium containing 15 muM kanamycin (negative selection), while in the positive selection strategy, shoots were selected on 2.5 g/l mannose supplemented with 15 g/l sucrose. Transformation efficiencies based on PCR analysis of individual putative transformed shoots from independent lines relative to the initial numbers of leaf explants tested were 5.6% for kanamycin/nptII and 6.8% for mannose/pmi selection, respectively. Southern blot analysis on six randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene in each, and five randomly chosen lines identified to contain the pmi transgene by PCR showed positive hybridisation to a pmi DNA probe. The positive (mannose/pmi) and the negative (kanamycin) selection protocols used in this study have greatly improved transformation efficiency in almond, which were confirmed with PCR and Southern blot. This study also demonstrates that in almond the mannose/pmi selection protocol is appropriate and can result in higher transformation efficiencies over that of kanamycin/nptII selection protocols.

  17. An efficient regeneration protocol for Agrobacterium-mediated transformation of melon (Cucumis melo L.).

    PubMed

    Zhang, H J; Gao, P; Wang, X Z; Luan, F S

    2014-01-08

    An efficient selection and plant regeneration protocol for Agrobacterium-mediated transformation, using cotyledon node zone-stem connection region of melon, has been developed. The new Agrobacterium-mediated transformation methodology, independent of organ culture, used the entire germinated seed as explants. The transformation system was maximized to maintain the integrity of melon itself, thus avoiding the limitations of traditional tissue culture methods. The transformation was carried out under a non-sterile environment. The incorporation of a selectable marker (neomycin phosphotransferase II) into the genome of transgenic plants was confirmed by PCR and Southern blot analyses. The transformation frequency based on the PCR was 13%. Transgenic melon plants were usually detected by PCR in less than 1 month after Agrobacterium inoculation, and seeds could be harvested in 3 months. The growth characteristics and morphology of the transgenic plants were identical to the untransformed wild-type plants. This method would be beneficial for facilitating the characteristics of gene functions and for boosting the manipulation of melon transformation for commercial purposes.

  18. Role of bacterial virulence proteins in Agrobacterium-mediated transformation of Aspergillus awamori.

    PubMed

    Michielse, C B; Ram, A F J; Hooykaas, P J J; Hondel, C A M J J van den

    2004-05-01

    The Agrobacterium-mediated transformation of Aspergillus awamori was optimized using defined co-cultivation conditions, which resulted in a reproducible and efficient transformation system. Optimal co-cultivation conditions were used to study the role of Agrobacterium tumefaciens virulence proteins in T-DNA transfer. This study revealed that inactivation of either of the regulatory proteins (VirA, VirG), any of the transport pore proteins (VirB), proteins involved in generation of the T-strand (VirD, VirC) or T-strand protection and targeting (VirE2) abolishes or severely reduces the formation of transformants. The results indicate that the Agrobacterium-mediated transformation of A. awamori requires an intact T-DNA machinery for efficient transformation; however, the plant host range factors, like VirE3, VirH, and VirF, are not important.

  19. Strategies to improve low copy transgenic events in Agrobacterium-mediated transformation of maize.

    PubMed

    Sivamani, Elumalai; Li, Xianggan; Nalapalli, Samson; Barron, Yoshimi; Prairie, Anna; Bradley, David; Doyle, Michele; Que, Qiudeng

    2015-12-01

    Transgenic plants containing low copy transgene insertion free of vector backbone are highly desired for many biotechnological applications. We have investigated two different strategies for increasing the percentage of low copy events in Agrobacterium-mediated transformation experiments in maize. One of the strategies is to use a binary vector with two separate T-DNAs, one T-DNA containing an intact E.coli manA gene encoding phosphomannose isomerase (PMI) as selectable marker gene cassette and another T-DNA containing an RNAi cassette of PMI sequences. By using this strategy, low copy transgenic events containing the transgenes were increased from 43 to 60 % in maize. An alternate strategy is using selectable marker gene cassettes containing regulatory or coding sequences derived from essential plant genes such as 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) or MADS box transcription factor. In this paper we demonstrate that higher percentage of low copy transgenic events can be obtained in Agrobacterium-mediated maize transformation experiments using both strategies. We propose that the above two strategies can be used independently or in combination to increase transgenic events that contain low copy transgene insertion in Agrobacterium-mediated transformation experiments.

  20. Agrobacterium-mediated genetic transformation of pineapple (Ananas comosus L., Merr.).

    PubMed

    Mhatre, Minal

    2013-01-01

    Pineapple (Ananas comosus L., Merr.) is a commercially important crop, grown in the tropical and subtropical regions. However, the crop is faced with postharvest damage and poor varietal and nutritional improvement. Being a vegetatively propagated crop, conventional breeding programs take longer time for genetic improvement, which may not necessarily successfully develop an improved cultivar. Hence, the genetic modification of pineapple is an alternative handy approach to improve pineapple. We have established an Agrobacterium-mediated transformation system using leaf bases from in vitro-grown pineapple plants. Being a monocot, acetosyringone is added to the culture medium for overnight growth of Agrobacterium and transformation to transfer a gene of interest MSI99 soybean ferritin. Leaf bases isolated from in vitro shoot cultures are treated with Agrobacterium suspension at two dilutions, 10× and 20×, for 30 min. Explants are subsequently blot dried and cultured on gelrite solidified hormone-free Pin1 medium for 2 days (cocultivation). Periodic transfer is first done to the regeneration medium (Pin1) containing cefotaxime for the suppression of Agrobacterium growth. The transformants are selected by culturing on Pin1 medium containing cefotaxime and kanamycin. Multiple shoots, regenerated in leaf bases, are further multiplied and individually rooted in the liquid RM medium amended with antibiotics to recover plants. Putative transformants are analyzed for transgene integration and expression using standard molecular biological methods of PCR, RT-PCR, and genomic Southern.

  1. AGROBACTERIUM-MEDIATED TRANSFORMATION IN THE GREEN ALGA HAEMATOCOCCUS PLUVIALIS (CHLOROPHYCEAE, VOLVOCALES)(1).

    PubMed

    Kathiresan, S; Chandrashekar, A; Ravishankar, G A; Sarada, R

    2009-06-01

    The first successful Agrobacterium-mediated transformation of the green alga Haematococcus pluvialis Flot. using the binary vectors hosting the genes coding for GUS (β-glucuronidase), GFP (green fluorescent protein), and hpt (hygromycin phosphotransferase) is reported here. Colonies resistant to hygromycin at 10 mg · L(-1) expressed β-glucuronidase. The greenish yellow fluorescence of GFP was observed when the hygromycin-resistant cells were viewed with a fluorescent microscope. PCR was used to successfully amplify fragments of the hpt (407 bp) and GUS (515 bp) genes from transformed cells, while Southern blots indicated the integration of the hygromycin gene into the genome of H. pluvialis. SEM indicated that the cell wall of H. pluvialis was altered on infection with Agrobacterium. The transformation achieved here by Agrobacterium does not need treatment with acetosyringone or the wounding of cells. A robust transformation method for this alga would pave the way for manipulation of many important pathways relevant to the food, pharmaceutical, and nutraceutical industries.

  2. [Agrobacterium-mediated transformation of LJAMP2 gene into 'Red Sun' kiwifruit and its molecular identification].

    PubMed

    Zhou, Yue; Zhao, Xupeng; Wu, Xiuhua; Zhang, Yanling; Zhang, Lin; Luo, Keming; Tang, Shaohu

    2014-06-01

    Bacterial canker caused by Pseudomonas syringae pv. Actinidiae is one of the most important diseases of kiwifruit (Actinidia chinensis) and leads to considerable yield losses. In order to obtain transgenic plants with resistance for 'Red Sun' kiwifruit to canker disease, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into 'Red Sun' kiwifruit through Agrobacterium-mediated transformation. After two days of co-cultivation with A. tumefaciens strain LBA4404 harboring 35S:LJAMP2, the transformed explants were transferred to the selection medium containing 25 mg/L kanamycin+3.0 mg/L BA+1.0 mg/L NAA. The regeneration efficiency of kanamycin-resistant shoots reached to 85%. All (100%) of kanamycin-resistant shoots rooted on half-strength MS medium supplemented with 0.8 mg/L IBA and a total of 40 regenerated plantlets were obtained. PCR and histochemical GUS activity analysis show that 23 of 40 lines (57.50%) were positive, suggesting that the LJAMP2 gene was integrated into the genome of 'Red Sun' kiwifruit. Taken together, we established an efficient genetic transformation method for 'Red Sun' kiwifruit using A. tumefaciens and the transformation frequency reached 5.11%. This protocol will be useful for the genetic breeding of 'Red Sun' kiwifruit for improvement of disease resistance.

  3. Transgenic sugar beet tolerant to imidazolinone obtained by Agrobacterium-mediated transformation.

    PubMed

    Kishchenko, E M; Komarnitskii, I K; Kuchuk, N V

    2011-01-01

    Sugar beet is highly sensitive to imidazolinone herbicides thus rotational restrictions exist. In order to develop imidazolinone tolerant sugar beets als gene from Arabidopsis thaliana encoding acetolactate synthase with S653N mutation was used for genetic transformation. Transgenic sugar beet plants were obtained by Agrobacterium-mediated transformation of aseptic seedlings using vacuum-infiltration. The efficiency of genetic transformation was 5.8%. RT-PCR analysis of obtained plants revealed accumulation of specific als transcript. The resistance to imidazolinone was proved for developed transgenic sugar beet plants in vitro and in greenhouse conditions after spraying with imazethapyr (Pursuit, BASF).

  4. Agrobacterium-mediated genetic transformation of Prunus salicina

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report Agrobacterium tumefaciens-mediated transformation from hypocotyls slices of two Prunus salicina varieties, 'Angeleno' and 'Larry Anne', using a modification of the technique previously described for P. domestica. Regeneration rates on thidiazuron (TDZ) and indole-3-butyric acid (IBA) supp...

  5. Development of Transgenic Papaya through Agrobacterium-Mediated Transformation

    PubMed Central

    Azad, Md. Abul Kalam; Rabbani, Md. Golam; Amin, Latifah; Sidik, Nik Marzuki

    2013-01-01

    Transgenic papaya plants were regenerated from hypocotyls and immature zygotic embryo after cocultivation with Agrobacterium tumefaciens LBA-4404 carrying a binary plasmid vector system containing neomycin phosphotransferase (nptII) gene as the selectable marker and β-glucuronidase (GUS) as the reporter gene. The explants were co-cultivated with Agrobacterium tumefaciens on regeneration medium containing 500 mg/L carbenicillin + 200 mg/L cefotaxime for one week. The cocultivated explants were transferred into the final selection medium containing 500 mg/L carbenicillin + 200 mg/L cefotaxime + 50 mg/L kanamycin for callus induction as well as plant regeneration. The callus derived from the hypocotyls of Carica papaya cv. Shahi showed the highest positive GUS activities compared to Carica papaya cv. Ranchi. The transformed callus grew vigorously and formed embryos followed by transgenic plantlets successfully. The result of this study showed that the hypocotyls of C. papaya cv. Shahi and C. papaya cv. Ranchi are better explants for genetic transformation compared to immature embryos. The transformed C. papaya cv. Shahi also showed the maximum number of plant regeneration compared to that of C. papaya cv. Ranchi. PMID:24066284

  6. High-efficiency Agrobacterium-mediated transformation of Norway spruce (Picea abies) and loblolly pine (Pinus taeda)

    NASA Technical Reports Server (NTRS)

    Wenck, A. R.; Quinn, M.; Whetten, R. W.; Pullman, G.; Sederoff, R.; Brown, C. S. (Principal Investigator)

    1999-01-01

    Agrobacterium-mediated gene transfer is the method of choice for many plant biotechnology laboratories; however, large-scale use of this organism in conifer transformation has been limited by difficult propagation of explant material, selection efficiencies and low transformation frequency. We have analyzed co-cultivation conditions and different disarmed strains of Agrobacterium to improve transformation. Additional copies of virulence genes were added to three common disarmed strains. These extra virulence genes included either a constitutively active virG or extra copies of virG and virB, both from pTiBo542. In experiments with Norway spruce, we increased transformation efficiencies 1000-fold from initial experiments where little or no transient expression was detected. Over 100 transformed lines expressing the marker gene beta-glucuronidase (GUS) were generated from rapidly dividing embryogenic suspension-cultured cells co-cultivated with Agrobacterium. GUS activity was used to monitor transient expression and to further test lines selected on kanamycin-containing medium. In loblolly pine, transient expression increased 10-fold utilizing modified Agrobacterium strains. Agrobacterium-mediated gene transfer is a useful technique for large-scale generation of transgenic Norway spruce and may prove useful for other conifer species.

  7. Agrobacterium-mediated transformation of Euphorbia tirucalli callus.

    PubMed

    Uchida, Hidenobu; Yamashita, Hirofumi; Anai, Toyoaki; Muranaka, Toshiya; Ohyama, Kanji

    2010-01-01

    In order to establish a basis for transformation technology in the petroleum plant Euphorbia tirucalli, the callus of the plant was infected with Agrobacterium, washed with distilled water, sterilized with distilled water containing 100 mg/l of carbenicillin, selected on solidified B5 medium containing 13 mg/l of G418 and 100 mg/l of carbenicillin, and then on solidified B5 medium containing 25 mg/l of G418 and 100 mg/l of carbenicillin for the transgenic calli, and then the callus lines were subcultured successively on solidified B5 medium containing 50 mg/l of G418. We performed PCR analysis of sterilized G418-resistant callus line DNA and concluded that the gene introduced was integrated into the callus genome.

  8. High frequency regeneration via direct somatic embryogenesis and efficient Agrobacterium- mediated genetic transformation of tobacco

    PubMed Central

    Pathi, Krishna Mohan; Tula, Suresh; Tuteja, Narendra

    2013-01-01

    A direct somatic embryogenesis protocol was developed for four cultivars of Nicotiana species, by using leaf disc as an explant. Direct somatic embryogenesis of Nicotiana by using BAP and IAA has not been investigated so far. This method does not require formation of callus tissues which leads to somaclonal variations. The frequency of somatic embryogenesis was strongly influenced by the plant growth hormones. The somatic embryos developing directly from explant tissue were noticed after 6 d of culture. Somatic embryogenesis of a high frequency (87–96%) was observed in cultures of the all four genotypes (Nicotiana tabacum, N. benthamiyana, N. xanthi, N. t cv petihavana). The results showed that the best medium for direct somatic embryogenesis was MS supplemented with 2.5 mg/l, 0.2 mg/l IAA and 2% sucrose. Subculture of somatic embryos onto hormone free MS medium resulted in their conversion into plants for all genotypes. About 95% of the regenerated somatic embryos germinated into complete plantlets. The plants showed morphological and growth characteristics similar to those of seed-derived plants. Explants were transformed using Agrobacterium tumifacious LBA4404 plasmid pCAMBIA1301 harboring the GUS gene. The regenerated transgenic plants were confirmed by PCR analysis and histochemical GUS assay. The transformation efficiency obtained by using the Agrobacterium- mediated transformation was more than 95%. This method takes 6 wk to accomplish complete transgenic plants through direct somatic embryogenesis. The transgenic plantlets were acclimatized successfully with 98% survival in greenhouse and they showed normal morphological characteristics and were fertile. The regeneration and transformation method described herein is very simple, highly efficient and fast for the introduction of any foreign gene directly in tobacco through direct somatic embryogenesis. PMID:23518589

  9. An embryogenic suspension cell culture system for Agrobacterium-mediated transformation of citrus.

    PubMed

    Dutt, M; Grosser, J W

    2010-11-01

    A method for the genetic transformation of several citrus cultivars is described, including cultivars observed to be recalcitrant to conventional epicotyl-mediated transformation. Embryogenic cell suspension cultures, established from unfertilized ovules were used as target tissues for Agrobacterium-mediated transformation. Several modifications were made to the culture environment to investigate factors required for efficient transfer of the T-DNA and the subsequent regeneration of transgenic citrus plants. It was determined that co-cultivation of citrus cells and Agrobacterium in EME medium supplemented with maltose (EME-M) and 100 μM acetosyringone for 5 days at 25°C was optimum for transformation of each of the citrus cultivars. Efficient selection was obtained and escapes were prevented when the antibiotic hygromycin B was used as a selection antibiotic following transformation with an Agrobacterium strain containing hptII in the T-DNA region. Transgenic embryo regeneration and development was enhanced in medium that contained a liquid overlay consisting of a 1:2 mixture of 0.6 M BH3 and 0.15 M EME-M media. PCR and Southern blot analyses confirmed the presence of the T-DNA and the stable integration into the genome of regenerated plants, while RT-PCR demonstrated variable amounts of RNA being transcribed in different transgenic lines. This protocol can create an avenue for insertion of useful traits into any polyembryonic citrus cultivar that can be established as embryogenic cell suspension cultures, including popular specialty mandarins and seedless cultivars.

  10. Agrobacterium-mediated large-scale transformation of wheat (Triticum aestivum L.) using glyphosate selection.

    PubMed

    Hu, T; Metz, S; Chay, C; Zhou, H P; Biest, N; Chen, G; Cheng, M; Feng, X; Radionenko, M; Lu, F; Fry, J

    2003-06-01

    An Agrobacterium-mediated transformation system with glyphosate selection has been developed for the large-scale production of transgenic plants. The system uses 4-day precultured immature embryos as explants. A total of 30 vectors containing the 5-enol-pyruvylshikimate-3-phosphate synthase gene from Agrobacterium strain CP4 (aroA:CP4), which confers resistance to glyphosate, were introduced into wheat using this system. The aroA:CP4 gene served two roles in this study-selectable marker and gene of interest. More than 3,000 transgenic events were produced with an average transformation efficiency of 4.4%. The entire process from isolation of immature embryos to production of transgenic plantlets was 50-80 days. Transgenic events were evaluated over several generations based on genetic, agronomic and molecular criteria. Forty-six percent of the transgenic events fit a 3:1 segregation ratio. Molecular analysis confirmed that four of six lead transgenic events selected from Agrobacterium transformation contained a single insert and a single copy of the transgene. Stable expression of theAROA:CP4 gene was confirmed by ELISA through nine generations. A comparison of Agrobacterium-mediated transformation to a particle bombardment system demonstrated that the Agrobacterium system is reproducible, has a higher transformation efficiency with glyphosate selection and produces higher quality transgenic events in wheat. One of the lead events from this study, no. 33391, has been identified as a Roundup Ready wheat commercial candidate.

  11. Efficient gene targeting in Penicillium chrysogenum using novel Agrobacterium-mediated transformation approaches.

    PubMed

    de Boer, Paulo; Bronkhof, Jurian; Dukiќ, Karolina; Kerkman, Richard; Touw, Hesselien; van den Berg, Marco; Offringa, Remko

    2013-12-01

    The industrial production of β-lactam antibiotics by Penicillium chrysogenum has increased tremendously over the last decades, however, further optimization via classical strain and process improvement has reached its limits. The availability of the genome sequence provides new opportunities for directed strain improvement, but this requires the establishment of an efficient gene targeting (GT) system. Recently, mutations affecting the non-homologous end joining (NHEJ) pathway were shown to increase GT efficiencies following PEG-mediated DNA transfer in P. chrysogenum from 1% to 50%. Apart from direct DNA transfer many fungi can efficiently be transformed using the T-DNA transfer system of the soil bacterium Agrobacterium tumefaciens, however, for P. chrysogenum no robust system for Agrobacterium-mediated transformation was available. We obtained efficient AMT of P. chrysogenum spores with the nourseothricin acetyltransferase gene as selection marker, and using this system we investigated if AMT in a NHEJ mutant background could further enhance GT efficiencies. In general, AMT resulted in higher GT efficiencies than direct DNA transfer, although the final frequencies depended on the Agrobacterium strain and plasmid backbone used. Providing overlapping and complementing fragments on two different plasmid backbones via the same Agrobacterium host was shown to be most effective. This so-called split-marker or bi-partite method resulted in highly efficient GT (>97%) almost exclusively without additional ectopic T-DNA insertions. As this method provides for an efficient GT method independent of protoplasts, it can be applied to other fungi for which no protoplasts can be generated or for which protoplast transformation leads to varying results.

  12. Factors influencing Agrobacterium-mediated embryogenic callus transformation of Valencia sweet orange (Citrus sinensis) containing the pTA29-barnase gene.

    PubMed

    Li, D D; Shi, W; Deng, X X

    2003-12-01

    Valencia sweet orange (Citrus sinensis (L.) Osbeck) calluses were used as explants to develop a new transformation system for citrus mediated by Agrobacterium tumefaciens. Factors affecting Agrobacterium-mediated transformation efficiency included mode of pre-cultivation, temperature of cocultivation and presence of acetosyringone (AS). The highest transformation efficiency was obtained with a 4-day pre-cultivation period in liquid medium. Transformation efficiency was higher when cocultivation was performed for 3 days at 19 degrees C than at 23 or 28 degrees C. Almost no resistant callus was obtained if the cocultivation medium lacked AS. The transformation procedure yielded transgenic Valencia plants containing the pTA29-barnase gene, as verified by PCR amplification and confirmed by Southern blotting. Because male sterility is a common factor leading to seedlessness in citrus cultivars with parthenocarpic characteristics, production of seedless citrus genotypes by Agrobacterium-mediated genetic transformation is a promising alternative to conventional breeding methods.

  13. Agrobacterium-mediated transformation of apricot (Prunus armeniaca L.) leaf explants.

    PubMed

    Petri, César; Wang, Hong; Alburquerque, Nuria; Faize, Mohamed; Burgos, Lorenzo

    2008-08-01

    A protocol for Agrobacterium-mediated stable transformation for scored, whole leaf explants of the apricot (Prunus armeniaca) cultivar Helena was developed. Regenerated shoots were selected using a two-step increased concentrations of paromomycin sulphate. Different factors affecting survival of transformed buds, including possible toxicity of green fluorescent protein (GFP) and time of exposure to high cytokine concentration in the regeneration medium, were examined. Transformation efficiency, based on PCR analysis of individual putative transformed shoots from independent lines was 5.6%, when optimal conditions for bud survival were provided. Southern blot analysis on four randomly chosen PCR-positive shoots confirmed the presence of the nptII transgene. This is the first time that stable transformation of an apricot cultivar is reported and constitutes also one of the few reports on the transformation of Prunus cultivars.

  14. A high-efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.).

    PubMed

    Ozawa, Kenjirou

    2012-01-01

    Agrobacterium-mediated transformation of rice has been routinely performed according to the protocol reported by Hiei et al. (Plant J. 6:271-282, 1994). However, several elite japonica and many indica varieties cannot be efficiently transformed by Agrobacterium system. Also a large number of transformants are required to generate T-DNA insertion and FOX libraries as well as gene-targeting studies. To overcome these challenges, we established a high-efficiency transformation system in rice by cocultivating rice calli with Agrobacterium on filter papers moistened with enriched liquid media instead of using solid media (Ozawa, Plant Sci. 176:522-527, 2009; Ozawa and Takaiwa, Plant Sci. 179:333-337, 2010). In this system, the transformation efficiency of the calli is almost 100% in many varieties.

  15. A simple and efficient Agrobacterium-mediated procedure for transformation of tomato.

    PubMed

    Sharma, Manoj K; Solanke, Amolkumar U; Jani, Dewal; Singh, Yogendra; Sharma, Arun K

    2009-09-01

    We describe a highly efficient and reproducible Agrobacterium-mediated transformation protocol applicable to several varieties of tomato (Solanum lycopersicum, earlier known as Lycopersicum esculentum). Conditions such as co-cultivation period, bacterial concentration, concentration of benzyl amino purine (BAP), zeatin and indole acetic acid (IAA) were optimized. Co-cultivation of explants with a bacterial concentration of 108 cells/ml for three days on 2 mg/l BAP, followed by regeneration on a medium containing 1 mg/ml zeatin resulted in a transformation frequency of 41.4%. Transformation of tomato plants was confirmed by Southern blot analysis and beta-glucuronidase (GUS) assay. The protocol developed showed very high efficiency of transformation for tomato varieties Pusa Ruby, Arka Vikas and Sioux. The optimized transformation procedure is simple, efficient and does not require tobacco, Petunia, tomato suspension feeder layer or acetosyringone.

  16. Agrobacterium-mediated transformation of Malus robusta with tomato iron transporter gene.

    PubMed

    Qu, Shen-Chun; Huang, Xiao-De; Zhang, Zhen; Yao, Quan-Hong; Tao, Jian-Min; Qiao, Yu-Shan; Zhang, Jun-Yi

    2005-06-01

    The tomato iron transporter gene (LeIRT2) was introduced to Malus robusta Rehd. via Agrobacterium-mediated transformation to produce iron-deficiency tolerant apple rootstock. A total of 19 putative transformants were obtained, 11 of which were verified by PCR amplification to carry a fragment of the transgene. Among them, nine were confirmed to carry the transgene by Southern blot analysis with one to three copies of the transgene integrated into the plant genome. Two transgenic plants, one carrying one copy and the other three copies of the transgene, were hydroponically cultured to test their tolerance to iron-deficiency, which was found only in the transgenic plant with a single copy, which weighted 21%-4% greater than those of the control plants.

  17. Agrobacterium-mediated transformation of the endophytic fungus Acremonium implicatum associated with Brachiaria grasses.

    PubMed

    Abello, Javier; Kelemu, Segenet; García, Celsa

    2008-03-01

    Acremonium implicatum is a seed-transmitted endophytic fungus that forms symbiotic associations with the economically significant tropical forage grasses, Brachiaria species. To take advantage of the endophyte's plant protective properties, we developed an efficient Agrobacterium-mediated transformation system for Acremonium implicatum, using green fluorescent protein (GFP) expression and vector pSK1019 (trpC promoter) or pCAMBIA1300 (CaMV35S promoter). We found that transformation efficiency doubled for both mycelial and conidial transformation as the co-cultivation period for Agrobacterium tumefaciens and Acremonium implicatum was increased from 48 to 72h. Significantly, optimal results were obtained for either mycelial or conidial transformation with Agrobacterium tumefaciens strain AGL-1 and vector pSK1019 under the control of the trpC promoter. However, mycelial transformation consistently generated a significantly higher number of transformants than did conidial transformation. The mitotic stability of the transferred DNA was confirmed by growing ten transformants in liquid and agar media for six generations. In all cases, resistance to the selection pressure (hygromycin B) was maintained. Fluorescence emission was retained by the transformants and also expressed in Brachiaria tissues from plants inoculated with GFP-transformed A. implicatum. This technology will help in the transfer and expression of agronomically important genes in host plants.

  18. Stable Agrobacterium-mediated transformation of maritime pine based on kanamycin selection.

    PubMed

    Alvarez, José M; Ordás, Ricardo J

    2013-01-01

    An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII) as a selectable marker gene and β -glucuronidase (uidA) as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL(-1) kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass) was achieved when a vigorously growing embryonal mass (embryogenic line L01) was cocultivated with Agrobacterium strain AGL1 at the optical density (OD(600 nm)) of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth.

  19. Stable Agrobacterium-Mediated Transformation of Maritime Pine Based on Kanamycin Selection

    PubMed Central

    Alvarez, José M.; Ordás, Ricardo J.

    2013-01-01

    An efficient transformation protocol based on kanamycin selection was developed for Agrobacterium-mediated transformation of maritime pine embryonal masses. The binary vector pBINUbiGUSint, which contained neomycin phosphotransferase II (nptII) as a selectable marker gene and β-glucuronidase (uidA) as a reporter gene, was used for transformation studies. Different factors, such as embryogenic line, bacterial strain, bacterial concentration, and coculture duration, were examined and optimized. For selection of transformants, 15 mgL−1 kanamycin was used. The highest transformation efficiency (11.4 events per gram of fresh mass) was achieved when a vigorously growing embryonal mass (embryogenic line L01) was cocultivated with Agrobacterium strain AGL1 at the optical density (OD600 nm) of 0.3 for 72 h. Evidence of the stable transgene integration was obtained by polymerase chain reaction for the nptII and uidA genes and expression of the uidA gene. Maturation capacity of the transgenic lines was negatively affected by the transformation process. Induction of axillary shoots by preculturing the embryos with benzyladenine allowed overcoming the low maturation rates of some transformed lines. The transgenic embryos were germinated and the axillar shoots were rooted. Transgenic plants were transferred to potting substrate showing normal growth. PMID:24376383

  20. Optimization of factors affecting Agrobacterium-mediated transformation of Micro-Tom tomatoes.

    PubMed

    Guo, M; Zhang, Y L; Meng, Z J; Jiang, J

    2012-03-16

    Micro-Tom is the smallest known variety of tomatoes. An orthogonal experimental design L(16) (4(5)) was used to optimize Agrobacterium-mediated transformation of cotyledon explants of Lycopersicon esculentum cv. Micro-Tom. Four parameters were investigated to determine their effect on transformation frequency: the concentration of bacterial suspension, time of dip in bacterial suspension, co-cultivation time, and concentration of carbenicillin. We also examined the effect of these parameters on contamination rate, necrosis rate, mortality, cut-surface browning rate, and undamaged explant rate. Both the bacterial and carbenicillin concentrations had a significant influence on the rate of infected explants. The time of co-cultivation also had a significant influence on the transformation parameters. The optimal transformation protocol consisted of an Agrobacterium suspension of 0.5 × 10(8) cells/mL (OD(600) = 0.5) and an infection time of 5 min, one day of co-cultivation and 500 mg/L carbenicillin. Under these conditions, the transformation efficiency of the shoots reached 5.1%; the mean transformation frequency was 3.9% (N = 838).

  1. Deletion of host histone acetyltransferases and deacetylases strongly affects Agrobacterium-mediated transformation of Saccharomyces cerevisiae.

    PubMed

    Soltani, Jalal; van Heusden, Gerard Paul H; Hooykaas, Paul J J

    2009-09-01

    Agrobacterium tumefaciens is a plant pathogen that genetically transforms plant cells by transferring a part of its Ti-plasmid, the T-strand, to the host cell. Under laboratory conditions, it can also transform cells from many different nonplant organisms, including the yeast Saccharomyces cerevisiae. Collections of S. cerevisiae strains have been developed with systematic deletion of all coding sequences. Here, we used these collections to identify genes involved in the Agrobacterium-mediated transformation (AMT) of S. cerevisiae. We found that deletion of genes (GCN5, NGG1, YAF9 and EAF7) encoding subunits of the SAGA, SLIK, ADA and NuA4 histone acetyltransferase complexes highly increased the efficiency of AMT, while deletion of genes (HDA2, HDA3 and HST4) encoding subunits of histone deacetylase complexes decreased AMT. These effects are specific for AMT as the efficiency of chemical (lithium acetate) transformation was not or only slightly affected by these deletions. Our data are consistent with a positive role of host histone deacetylation in AMT.

  2. Agrobacterium-mediated transformation of oat (Avena sativa L.) cultivars via immature embryo and leaf explants.

    PubMed

    Gasparis, Sebastian; Bregier, Cezary; Orczyk, Waclaw; Nadolska-Orczyk, Anna

    2008-11-01

    This paper reports on the successful Agrobacterium-mediated transformation of oat, and on some factors influencing this process. In the first step of the experiments, three cultivars, two types of explant, and three combinations of strain/vectors, which were successfully used for transformation of other cereals were tested. Transgenic plants were obtained from the immature embryos of cvs. Bajka, Slawko and Akt and from leaf base explants of cv. Bajka after transformation with A. thumefaciens strain LBA4404(pTOK233). The highest transformation rate (12.3%) was obtained for immature embryos of cv. Bajka. About 79% of the selected plants proved to be transgenic; however, only 14.3% of the T(0) plants and 27.5% of the T(1) showed GUS expression. Cell competence of both types of explant differed in terms of their transformation ability and transgene expression. The next step of the study was to test the suitability for oat transformation of the pGreen binary vector combined with different selection cassettes: nptII or bar under the nos or 35S promoter. Transgenic plants were selected in combinations transformed with nos::nptII, 35S::nptII and nos::bar. The highest transformation efficiency (5.3%) was obtained for cv. Akt transformed with nos::nptII. A detailed analysis of the T(0) plants selected from a given callus line and their progeny revealed that they were the mixture of transgenic, chimeric-transgenic and non-transgenic individuals. Southern blot analysis of T(0) and T(1) showed simple integration pattern with the low copy number of the introduced transgenes.

  3. Improved Agrobacterium-mediated transformation and high efficiency of root formation from hypocotyl meristem of spring Brassica napus 'Precocity' cultivar.

    PubMed

    Liu, X X; Lang, S R; Su, L Q; Liu, X; Wang, X F

    2015-12-14

    Rape seed (Brassica napus L.) is one of the most important oil seed crops in the world. Genetic manipulation of rapeseed requires a suitable tissue culture system and an efficient method for plant regeneration, as well as an efficient transformation procedure. However, development of transgenic B. napus has been problematic, and current studies are limited to cultivated varieties. In this study, we report a protocol for regeneration of transgenic rape after Agrobacterium-mediated transformation of hypocotyls from the spring B. napus 'Precocity' cultivar. We analyzed the effects of plant growth regulators in the medium on regeneration. Additionally, factors affecting the transformation efficiency, including seedling age, Agrobacterium concentration, infection time, and co-cultivation time, were assessed by monitoring GUS expression. Results from these experiments revealed that transformation was optimized when the meristematic parts of the hypocotyls were taken from 8 day-old seedlings, cultured on Murashinge and Skoog basal media containing 0.1 mg/L 1-naphthaleneacetic acid and 2.5 mg/L 6-benzylaminopurine, and incubated in Agrobacterium suspension (OD600 = 0.5) for 3 to 5 min, followed by 2 days of co-cultivation. Integration of T-DNA into the plant genome was confirmed by polymerase chain reaction (PCR), b-glucuronidase histochemical staining, and quantitative real-time PCR. The protocols developed for regeneration, transformation, and rooting described in this study could help to accelerate the development of transgenic spring rape varieties with novel features.

  4. An improved Agrobacterium-mediated transformation system for the functional genetic analysis of Penicillium marneffei.

    PubMed

    Kummasook, Aksarakorn; Cooper, Chester R; Vanittanakom, Nongnuch

    2010-12-01

    We have developed an improved Agrobacterium-mediated transformation (AMT) system for the functional genetic analysis of Penicillium marneffei, a thermally dimorphic, human pathogenic fungus. Our AMT protocol included the use of conidia or pre-germinated conidia of P. marneffei as the host recipient for T-DNA from Agrobacterium tumefaciens and co-cultivation at 28°C for 36 hours. Bleomycin-resistant transformants were selected as yeast-like colonies following incubation at 37°C. The efficiency of transformation was approximately 123 ± 3.27 and 239 ± 13.12 transformants per plate when using 5 × 10(4) conidia and pre-germinated conidia as starting materials, respectively. Southern blot analysis demonstrated that 95% of transformants contained single copies of T-DNA. Inverse PCR was employed for identifying flanking sequences at the T-DNA insertion sites. Analysis of these sequences indicated that integration occurred as random recombination events. Among the mutants isolated were previously described stuA and gasC defective strains. These AMT-derived mutants possessed single T-DNA integrations within their particular coding sequences. In addition, other morphological and pigmentation mutants possessing a variety of gene-specific defects were isolated, including two mutants having T-DNA integrations within putative promoter regions. One of the latter integration events was accompanied by the deletion of the entire corresponding gene. Collectively, these results indicated that AMT could be used for large-scale, functional genetic analyses in P. marneffei. Such analyses can potentially facilitate the identification of those genetic elements related to morphogenesis, as well as pathogenesis in this medically important fungus.

  5. Agrobacterium-mediated genetic transformation of Coffea arabica (L.) is greatly enhanced by using established embryogenic callus cultures

    PubMed Central

    2011-01-01

    Background Following genome sequencing of crop plants, one of the main challenges today is determining the function of all the predicted genes. When gene validation approaches are used for woody species, the main obstacle is the low recovery rate of transgenic plants from elite or commercial cultivars. Embryogenic calli have frequently been the target tissue for transformation, but the difficulty in producing or maintaining embryogenic tissues is one of the main problems encountered in genetic transformation of many woody plants, including Coffea arabica. Results We identified the conditions required for successful long-term proliferation of embryogenic cultures in C. arabica and designed a highly efficient and reliable Agrobacterium tumefaciens-mediated transformation method based on these conditions. The transformation protocol with LBA1119 harboring pBin 35S GFP was established by evaluating the effect of different parameters on transformation efficiency by GFP detection. Using embryogenic callus cultures, co-cultivation with LBA1119 OD600 = 0.6 for five days at 20 °C enabled reproducible transformation. The maintenance conditions for the embryogenic callus cultures, particularly a high auxin to cytokinin ratio, the age of the culture (optimum for 7-10 months of proliferation) and the use of a yellow callus phenotype, were the most important factors for achieving highly efficient transformation (> 90%). At the histological level, successful transformation was related to the number of proembryogenic masses present. All the selected plants were proved to be transformed by PCR and Southern blot hybridization. Conclusion Most progress in increasing transformation efficiency in coffee has been achieved by optimizing the production conditions of embryogenic cultures used as target tissues for transformation. This is the first time that a strong positive effect of the age of the culture on transformation efficiency was demonstrated. Our results make Agrobacterium-mediated

  6. Agrobacterium-mediated genetic transformation of yam (Dioscorea rotundata): an important tool for functional study of genes and crop improvement

    PubMed Central

    Nyaboga, Evans; Tripathi, Jaindra N.; Manoharan, Rajesh; Tripathi, Leena

    2014-01-01

    Although genetic transformation of clonally propagated crops has been widely studied as a tool for crop improvement and as a vital part of the development of functional genomics resources, there has been no report of any existing Agrobacterium-mediated transformation of yam (Dioscorea spp.) with evidence of stable integration of T-DNA. Yam is an important crop in the tropics and subtropics providing food security and income to over 300 million people. However, yam production remains constrained by increasing levels of field and storage pests and diseases. A major constraint to the development of biotechnological approaches for yam improvement has been the lack of an efficient and robust transformation and regeneration system. In this study, we developed an Agrobacterium-mediated transformation of Dioscorea rotundata using axillary buds as explants. Two cultivars of D. rotundata were transformed using Agrobacterium tumefaciens harboring the binary vectors containing selectable marker and reporter genes. After selection with appropriate concentrations of antibiotic, shoots were developed on shoot induction and elongation medium. The elongated antibiotic-resistant shoots were subsequently rooted on medium supplemented with selection agent. Successful transformation was confirmed by polymerase chain reaction, Southern blot analysis, and reporter genes assay. Expression of gusA gene in transgenic plants was also verified by reverse transcription polymerase chain reaction analysis. Transformation efficiency varied from 9.4 to 18.2% depending on the cultivars, selectable marker genes, and the Agrobacterium strain used for transformation. It took 3–4 months from Agro-infection to regeneration of complete transgenic plant. Here we report an efficient, fast and reproducible protocol for Agrobacterium-mediated transformation of D. rotundata using axillary buds as explants, which provides a useful platform for future genetic engineering studies in this economically important

  7. Application of sonication in combination with vacuum infiltration enhances the Agrobacterium-mediated genetic transformation in Indian soybean cultivars.

    PubMed

    Arun, Muthukrishnan; Subramanyam, Kondeti; Mariashibu, Thankaraj Salammal; Theboral, Jeevaraj; Shivanandhan, Ganeshan; Manickavasagam, Markandan; Ganapathi, Andy

    2015-02-01

    Soybean is a recalcitrant crop to Agrobacterium-mediated genetic transformation. Development of highly efficient, reproducible, and genotype-independent transformation protocol is highly desirable for soybean genetic improvement. Hence, an improved Agrobacterium-mediated genetic transformation protocol has been developed for cultivar PK 416 by evaluating various parameters including Agrobacterium tumefaciens strains (LBA4404, EHA101, and EHA105 harboring pCAMBIA1304 plasmid), sonication duration, vacuum infiltration pressure, and vacuum duration using cotyledonary node explants of soybean prepared from 7-day-old seedlings. The transformed plants were successfully developed through direct organogenesis system. Transgene expression was assessed by GUS histochemical and gfp visual assays, and integration was analyzed by PCR and Southern blot hybridization. Among the different combinations and durations evaluated, a maximum transformation efficiency of 18.6 % was achieved when the cotyledonary node explants of cv. PK 416 were sonicated for 20 s and vacuum infiltered for 2 min at 250 mmHg in A. tumefaciens EHA105 suspension. The amenability of the standardized protocol was tested on four more soybean cultivars JS 90-41, Hara Soy, Co 1, and Co 2 in which all the cultivars responded favorably with transformation efficiency ranging from 13.3 to 16.6 %. The transformation protocol developed in the present study would be useful to transform diverse soybean cultivars with desirable traits.

  8. Mitochondrial Porin Isoform AtVDAC1 Regulates the Competence of Arabidopsis thaliana to Agrobacterium-Mediated Genetic Transformation

    PubMed Central

    Kwon, Tackmin

    2016-01-01

    The efficiency of Agrobacterium-mediated transformation in plants depends on the virulence of Agrobacterium strains, the plant tissue culture conditions, and the susceptibility of host plants. Understanding the molecular interactions between Agrobacterium and host plant cells is crucial when manipulating the susceptibility of recalcitrant crop plants and protecting orchard trees from crown gall disease. It was discovered that Arabidopsis voltage-dependent anion channel 1 (atvdac1) mutant has drastic effects on Agrobacterium-mediated tumorigenesis and growth developmental phenotypes, and that these effects are dependent on a Ws-0 genetic background. Genetic complementation of Arabidopsis vdac1 mutants and yeast porin1-deficient strain with members of the AtVDAC gene family revealed that AtVDAC1 is required for Agrobacterium-mediated transformation, and there is weak functional redundancy between AtVDAC1 and AtVDAC3, which is independent of porin activity. Furthermore, atvdac1 mutants were deficient in transient and stable transformation by Agrobacterium, suggesting that AtVDAC1 is involved in the early stages of Agrobacterium infection prior to transferred-DNA (T-DNA) integration. Transgenic plants overexpressing AtVDAC1 not only complemented the phenotypes of the atvdac1 mutant, but also showed high efficiency of transient T-DNA gene expression; however, the efficiency of stable transformation was not affected. Moreover, the effect of phytohormone treatment on competence to Agrobacterium was compromised in atvdac1 mutants. These data indicate that AtVDAC1 regulates the competence of Arabidopsis to Agrobacterium infection. PMID:27643450

  9. Transgenic plants from shoot apical meristems of Vitis vinifera L. "Thompson Seedless" via Agrobacterium-mediated transformation.

    PubMed

    Dutt, M; Li, Z T; Dhekney, S A; Gray, D J

    2007-12-01

    Shoot apical meristem explants of Vitis vinifera "Thompson Seedless" were used for Agrobacterium-mediated genetic transformation. It was determined that the meristems had to be subjected to a dark growth phase then wounded to obtain transgenic plants. Morphological and histological studies illustrated the role of wounding to expose apical meristem cells for transformation. A bifunctional egfp/nptII fusion gene was used to select kanamycin resistant plants that expressed green fluorescent protein (GFP). Kanamycin at a concentration of 16 mg L(-1) in selection medium resulted in recovery of non-chimeric transgenic plants that uniformly expressed GFP, whereas 8 mg L(-1) kanamycin allowed non-transgenic and/or chimeric plants to develop. Polymerase chain reaction (PCR) and Southern blot analyses confirmed the presence of transgenes and their stable integration into the genome of regenerated plants. Up to 1% of shoot tips produced stable transgenic cultures within 6 weeks of treatment, resulting in a total of 18 independent lines.

  10. Improvement of Agrobacterium-mediated transformation in Hi-II maize (Zea mays) using standard binary vectors.

    PubMed

    Vega, Juan M; Yu, Weichang; Kennon, Angela R; Chen, Xinlu; Zhang, Zhanyuan J

    2008-02-01

    High-frequency transformation of maize (Zea mays L.) using standard binary vectors is advantageous for functional genomics and other genetic engineering studies. Recent advances in Agrobacterium tumefaciens-mediated transformation of maize have made it possible for the public to transform maize using standard binary vectors without a need of the superbinary vector. While maize Hi-II has been a preferred maize genotype to use in various maize transformation efforts, there is still potential and need in further improving its transformation frequency. Here we report the enhanced Agrobacterium-mediated transformation of immature zygotic embryos of maize Hi-II using standard binary vectors. This improved transformation process employs low-salt media in combined use with antioxidant L-cysteine alone or L-cysteine and dithiothreitol (DTT) during the Agrobacterium infection stage. Three levels of N6 medium salts, 10, 50, and 100%, were tested. Both 10 and 50% salts were found to enhance the T-DNA transfer in Hi-II. Addition of DTT to the cocultivation medium also improves the T-DNA transformation. About 12% overall and the highest average of 18% transformation frequencies were achieved from a large number of experiments using immature embryos grown in various seasons. The enhanced transformation protocol established here will be advantageous for maize genetic engineering studies including transformation-based functional genomics.

  11. Efficient Agrobacterium-mediated transformation of commercial hybrid poplar Populus nigra L. x P. maximowiczii A. Henry.

    PubMed

    Yevtushenko, Dmytro P; Misra, Santosh

    2010-03-01

    Many economically important species of Populus, especially those in sections Aigeiros and Tacamahaca, remain recalcitrant to genetic transformation. In this study, a simple and reliable protocol was developed for the efficient Agrobacterium-mediated transformation of a difficult-to-transform, but commercially viable, hybrid poplar Populus nigra L. x P. maximowiczii A. Henry (NM6). A plant transformation vector designed to express the beta-glucuronidase (GUS) gene was used to detect transformation events at early stages of plant regeneration and to optimize parameters affecting poplar transformation. The use of zeatin riboside in shoot-induction medium, regeneration of shoots via indirect organogenesis, and early selection pressure were the major modifications that drastically improved the efficiency of poplar transformation and minimized the number of untransformed regenerants. Transgenic shoots were routinely obtained 4-10 weeks after co-culture with A. tumefaciens, with a greater than 90% rate of plant recovery. Stable transgene integration, ranging from a single insertion to ten copies per genome, was confirmed by Southern blot analysis. The mean transformation frequency was 36.3% and about two-thirds of the lines had 1-2 transgene copies. Among the explants, petioles and leaves had a higher transformation frequency than did stem segments. Growth characteristics and the morphology of transgenic poplar plants were identical to untransformed controls. These findings will accelerate the development of P. nigra x P. maximowiczii plants with novel traits, and may also be useful to improve transformation procedures for other Populus species.

  12. Highly Efficient Agrobacterium-Mediated Transformation of Wheat Via In Planta Inoculation

    NASA Astrophysics Data System (ADS)

    Risacher, Thierry; Craze, Melanie; Bowden, Sarah; Paul, Wyatt; Barsby, Tina

    This chapter details a reproducible method for the transformation of spring wheat using Agrobacterium tumefaciens via the direct inoculation of bacteria into immature seeds in planta as described in patent WO 00/63398(1. Transformation efficiencies from 1 to 30% have been obtained and average efficiencies of at least 5% are routinely achieved. Regenerated plants are phenotypically normal with 30-50% of transformation events carrying introduced genes at single insertion sites, a higher rate than is typically reported for transgenic plants produced using biolistic transformation methods.

  13. Development of a phosphomannose isomerase-based Agrobacterium-mediated transformation system for chickpea (Cicer arietinum L.).

    PubMed

    Patil, Gunvant; Deokar, Amit; Jain, P K; Thengane, R J; Srinivasan, R

    2009-11-01

    To develop an alternative genetic transformation system that is not dependent on an antibiotic selection strategy, the phosphomannose isomerase gene (pmi) system was evaluated for producing transgenic plants of chickpea (Cicer arietinum L.). A shoot morphogenesis protocol based on the thidiazuron (TDZ)-induced shoot morphogenesis system was combined with Agrobacterium-mediated transformation of the pmi gene and selection of transgenic plants on mannose. Embryo axis explants of chickpea cv. C-235 were grown on a TDZ-supplemented medium for shoot proliferation. Embryo axis explants from which the first and second flush of shoots were removed were transformed using Agrobacterium carrying the pmi gene, and emerging shoots were allowed to regenerate on a zeatin-supplemented medium with an initial selection pressure of 20 g l(-1) mannose. Rooting was induced in the selected shoots on an indole-3-butyric acid (IBA)-supplemented medium with a selection pressure of 15 g l(-1) mannose. PCR with marker gene-specific primers and chlorophenol red (CPR) assay of the shoots indicated that shoots had been transformed. RT-PCR and Southern analysis of selected regenerated plants further confirmed integration of the transgene into the chickpea genome. These positive results suggest that the pmi/mannose selection system can be used to produce transgenic plants of chickpea that are free from antibiotic resistance marker genes.

  14. Agrobacterium-mediated genetic transformation of the desiccation tolerant resurrection plant Ramonda myconi (L.) Rchb.

    PubMed

    Tóth, Sándor; Kiss, Csaba; Scott, Peter; Kovács, Gabriella; Sorvari, Seppo; Toldi, Ottó

    2006-05-01

    In this paper we describe the first procedure for Agrobacterium tumefaciens-mediated genetic transformation of the desiccation tolerant plant Ramonda myconi (L.) Rchb. Previously, we reported the establishment of a reliable and effective tissue culture system based on the integrated optimisation of antioxidant and growth regulator composition and the stabilisation of the pH of the culture media by means of a potassium phosphate buffer. This efficient plant regeneration via callus phase provided a basis for the optimisation of the genetic transformation in R. myconi. For gene delivery, both a standard (method A) and a modified protocol (method B) have been applied. Since the latter has previously resulted in successful transformation of another resurrection plant, Craterostigma plantagineum, an identical protocol was utilized in transformation of R. myconi, as this method may prove general for dicotyledonous resurrection plants. On this basis, physical and biochemical key variables in transformation were evaluated such as mechanical microwounding of plant explants and in vitro preinduction of vir genes. While the physical enhancement of bacterial penetration was proved to be essential for successful genetic transformation of R. myconi, an additional two-fold increase in the transformation frequency was obtained when the above physical and biochemical treatments were applied in combination. All R0 and R1 transgenic plants were fertile, and no morphological abnormalities were observed on the whole-plant level.

  15. Refined glufosinate selection in Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill].

    PubMed

    Zeng, P; Vadnais, D A; Zhang, Z; Polacco, J C

    2004-02-01

    Modern genetic analysis and manipulation of soybean ( Glycine max) depend heavily on an efficient and dependable transformation process, especially in public genotypes from which expressed sequence tag (EST), bacterial artificial chromosome and microarray data have been derived. Williams 82 is the subject of EST and functional genomics analyses. However, it has not previously been transformed successfully using either somatic embryogenesis-based or cotyledonary-node transformation methods, the two predominant soybean transformation systems. An advance has recently been made in using antioxidants to enhance Agrobacterium infection of soybean. Nonetheless, an undesirable effect of using these antioxidants is the compromised recovery of transgenic soybean when combined with the use of the herbicide glufosinate as a selective agent. Therefore, we optimized both Agrobacterium infection and glufosinate selection in the presence of L-cysteine for Williams 82. We have recovered transgenic lines of this genotype with an enhanced transformation efficiency using this herbicide selection system.

  16. Genetic transformation of Metroxylon sagu (Rottb.) cultures via Agrobacterium-mediated and particle bombardment.

    PubMed

    Ibrahim, Evra Raunie; Hossain, Md Anowar; Roslan, Hairul Azman

    2014-01-01

    Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance.

  17. Genetic Transformation of Metroxylon sagu (Rottb.) Cultures via Agrobacterium-Mediated and Particle Bombardment

    PubMed Central

    Ibrahim, Evra Raunie

    2014-01-01

    Sago palm (Metroxylon sagu) is a perennial plant native to Southeast Asia and exploited mainly for the starch content in its trunk. Genetic improvement of sago palm is extremely slow when compared to other annual starch crops. Urgent attention is needed to improve the sago palm planting material and can be achieved through nonconventional methods. We have previously developed a tissue culture method for sago palm, which is used to provide the planting materials and to develop a genetic transformation procedure. Here, we report the genetic transformation of sago embryonic callus derived from suspension culture using Agrobacterium tumefaciens and gene gun systems. The transformed embryoids cells were selected against Basta (concentration 10 to 30 mg/L). Evidence of foreign genes integration and function of the bar and gus genes were verified via gene specific PCR amplification, gus staining, and dot blot analysis. This study showed that the embryogenic callus was the most suitable material for transformation as compared to the fine callus, embryoid stage, and initiated shoots. The gene gun transformation showed higher transformation efficiency than the ones transformed using Agrobacterium when targets were bombarded once or twice using 280 psi of helium pressure at 6 to 8 cm distance. PMID:25295258

  18. Agrobacterium-mediated transformation of safflower and the efficient recovery of transgenic plants via grafting

    PubMed Central

    2011-01-01

    Background Safflower (Carthamus tinctorius L.) is a difficult crop to genetically transform being susceptible to hyperhydration and poor in vitro root formation. In addition to traditional uses safflower has recently emerged as a broadacre platform for the production of transgenic products including modified oils and pharmaceutically active proteins. Despite commercial activities based on the genetic modification of safflower, there is no method available in the public domain describing the transformation of safflower that generates transformed T1 progeny. Results An efficient and reproducible protocol has been developed with a transformation efficiency of 4.8% and 3.1% for S-317 (high oleic acid content) and WT (high linoleic acid content) genotypes respectively. An improved safflower transformation T-DNA vector was developed, including a secreted GFP to allow non-destructive assessment of transgenic shoots. Hyperhydration and necrosis of Agrobacterium-infected cotyledons was effectively controlled by using iota-carrageenan, L-cysteine and ascorbic acid. To overcome poor in vitro root formation for the first time a grafting method was developed for safflower in which ~50% of transgenic shoots develop into mature plants bearing viable transgenic T1 seed. The integration and expression of secreted GFP and hygromycin genes were confirmed by PCR, Southern and Western blot analysis. Southern blot analysis in nine independent lines indicated that 1-7 transgenes were inserted per line and T1 progeny displayed Mendelian inheritance. Conclusions This protocol demonstrates significant improvements in both the efficiency and ease of use over existing safflower transformation protocols. This is the first complete method of genetic transformation of safflower that generates stably-transformed plants and progeny, allowing this crop to benefit from modern molecular applications. PMID:21595986

  19. A high-throughput Agrobacterium-mediated transformation system for the grass model species Brachypodium distachyon L.

    PubMed

    Păcurar, Daniel Ioan; Thordal-Christensen, Hans; Nielsen, Klaus Kristian; Lenk, Ingo

    2008-10-01

    In the ongoing process of developing Brachypodium distachyon as a model plant for temperate cereals and forage grasses, we have developed a high-throughput Agrobacterium-mediated transformation system for a diploid accession. Embryogenic callus, derived from immature embryos of the accession BDR018, were transformed with Agrobacterium tumefaciens strain AGL1 carrying two T-DNA plasmids, pDM805 and pWBV-Ds-Ubi-bar-Ds. Transient and stable transformation efficiencies were optimised by varying the pre-cultivation period, which had a strong effect on stable transformation efficiency. On average 55% of 17-day-old calli co-inoculated with Agrobacterium regenerated stable transgenic plants. Stable transformation frequencies of up to 80%, which to our knowledge is the highest transformation efficiency reported in graminaceous species, were observed. In a study of 177 transgenic lines transformed with pDM805, all of the regenerated transgenic lines were resistant to BASTA, while the gusA gene was expressed in 88% of the transgenic lines. Southern blot analysis revealed that 35% of the tested plants had a single T-DNA integration. Segregation analysis performed on progenies of ten selected T(0) plants indicated simple Mendelian inheritance of the two transgenes. Furthermore, the presence of two selection marker genes, bar and hpt, on the T-DNA of pWBV-Ds-Ubi-bar-Ds allowed us to characterize the developed transformation protocol with respect to full-length integration rate. Even when not selected for, full-length integration occurred in 97% of the transformants when using bialaphos as selection agent.

  20. Agrobacterium-Mediated Stable Genetic Transformation of Populus angustifolia and Populus balsamifera

    PubMed Central

    Maheshwari, Priti; Kovalchuk, Igor

    2016-01-01

    The present study demonstrates Agrobacterium tumefaciens-mediated stable genetic transformation of two species of poplar – Populus angustifolia and Populus balsamifera. The binary vector pCAMBIA-Npro-long-Luc containing the luciferase reporter gene was used to transform stem internode and axillary bud explants. Putative transformants were regenerated on selection-free medium using our previously established in vitro regeneration method. Explant type, genotype, effect of pre-culture, Agrobacterium concentration, a time period of infection and varying periods of co-culture with bacteria were tested for the transformation frequency. The highest frequency of transformation was obtained with stem internode explants pre-cultured for 2 days, infected with Agrobacterium culture at the concentration of OD600 = 0.5 for 10 min and co-cultivated with Agrobacterium for 48 h. Out of the two genotypes tested, P. balsamifera exhibited a higher transformation rate in comparison to P. angustifolia. The primary transformants that exhibited luciferase activity in a bioluminescence assay under the CCD camera when subjected to polymerase chain reaction and Southern blot analysis revealed a stable single-copy integration of luc in their genomes. The reported protocol is highly reproducible and can be applied to other species of poplar; it will also be useful for future genetic engineering of one of the most important families of woody plants for sustainable development. PMID:27014319

  1. Production of purple-colored creeping bentgrass using maize transcription factor genes Pl and Lc through Agrobacterium-mediated transformation.

    PubMed

    Han, Yun-Jeong; Kim, Yong-Min; Lee, Jee-Yeon; Kim, Soo Jung; Cho, Kyu-Chang; Chandrasekhar, Thummala; Song, Pill-Soon; Woo, Young-Min; Kim, Jeong-Il

    2009-03-01

    Purple-colored transgenic creeping bentgrass (Agrostis stolonifera L.) plants were developed for ornamental purpose by means of Agrobacterium-mediated transformation. Embryogenic creeping bentgrass calli were transformed with the pCAMBIA 3301 vector harboring maize (Zea mays) flavonoid/anthocyanin biosynthetic pathway transcription factor genes, Lc (Leaf color) and Pl (Purple leaf), individually and in combination, and three types of putative transgenic plants (Lc, Pl, and Lc + Pl) were generated. Genomic integration and expression of the transgenes were confirmed by Southern and northern blot analyses, respectively. The transgenic creeping bentgrass plants expressing both Lc and Pl genes were entirely purple, whereas those expressing Pl alone had purple stems and those expressing Lc alone lacked purple pigmentation in adult plants. The anthocyanin content was estimated in all the three types of transgenic plant and correlated well with the degree of purple coloration observed. These results suggest that both Lc and Pl genes are necessary and sufficient to confer purple coloration to creeping bentgrass.

  2. Establishment of a high efficiency Agrobacterium-mediated transformation system of rice (Oryza sativa L.).

    PubMed

    Ozawa, Kenjirou

    2009-04-01

    Technologies for transformation of rice have been developed to meet the requirements of functional genomics in order to enable the production of transgenic rice plants with useful agricultural characters. However, many rice varieties are not efficiently transformed by Agrobacterium. We have succeeded in establishing a highly efficient transformation system in rice by co-cultivating rice calli with Agrobacterium on three filter papers moistened with enriched N6 or DKN media instead of using solid media. Rice calli immersed in Agrobacterium suspension (EHA101, Agrobacterium concentration of OD600=0.04) were co-cultured on three pieces of filter paper (9cm in diameter) moistened with 5.5mL of N6 or DKN liquid co-cultivation medium supplemented with 2,4-d (2mg/L), proline (10mM), casein hydrolysate (300mg/L), sucrose (30g/L), glucose (5g/L), l-cysteine (100mg/L) and acetosyringone (15mg/L) at 25°C for 3 days in the dark. Compared with the transformation efficiency of calli co-cultivated on solid media, transformation efficiency was increased by about fivefold by using the filter paper method for many varieties of rice, including those that previously yielded much poor transformation rates.

  3. Use of the GFP reporter as a vital marker for Agrobacterium-mediated transformation of sugar beet (Beta vulgaris L.).

    PubMed

    Zhang, C L; Chen, D F; McCormac, A C; Scott, N W; Elliott, M C; Slater, A

    2001-02-01

    Molecular approaches to sugar beet improvement will benefit from an efficient transformation procedure that does not rely upon exploitation of selectable marker genes such as those which confer antibiotic or herbicide resistance upon the transgenic plants. The expression of the green fluorescent protein (GFP) signal has been investigated during a program of research that was designed to address the need to increase the speed and efficiency of selection of sugar beet transformants. It was envisaged that the GFP reporter could be used initially as a supplement to current selection regimes in order to help eliminate "escapes" and perhaps eventually as a replacement marker in order to avoid the public disquiet associated with antibiotic/herbicide-resistance genes in field-released crops. The sgfp-S65T gene has been modified to have a plant-compatible codon usage, and a serine to threonine mutation at position 65 for enhanced fluorescence under blue light. This gene, under the control of the CaMV 35S promoter, was introduced into sugar beet via Agrobacterium-mediated transformation. Early gene expression in cocultivated sugar beet cultures was signified by green fluorescence several days after cocultivation. Stably transformed calli, which showed green fluorescence at a range of densities, were obtained at frequencies of 3-11% after transferring the inoculated cultures to selection media. Cocultivated shoot explants or embryogenic calli were regularly monitored under the microscope with blue light when they were transferred to media without selective agents. Green fluorescent shoots were obtained at frequencies of 2-5%. It was concluded that the sgfp-S65T gene can be used as a vital marker for noninvasive screening of cells and shoots for transformation, and that it has potential for the development of selectable marker-free transgenic sugar beet.

  4. Generation of Marker- and/or Backbone-Free Transgenic Wheat Plants via Agrobacterium-Mediated Transformation

    PubMed Central

    Wang, Gen-Ping; Yu, Xiu-Dao; Sun, Yong-Wei; Jones, Huw D.; Xia, Lan-Qin

    2016-01-01

    Horizontal transfer of antibiotic resistance genes to animals and vertical transfer of herbicide resistance genes to the weedy relatives are perceived as major biosafety concerns in genetically modified (GM) crops. In this study, five novel vectors which used gusA and bar as a reporter gene and a selection marker gene, respectively, were constructed based on the pCLEAN dual binary vector system. Among these vectors, 1G7B and 5G7B carried two T-DNAs located on two respective plasmids with 5G7B possessing an additional virGwt gene. 5LBTG154 and 5TGTB154 carried two T-DNAs in the target plasmid with either one or double right borders, and 5BTG154 carried the selectable marker gene on the backbone outside of the T-DNA left border in the target plasmid. In addition, 5BTG154, 5LBTG154, and 5TGTB154 used pAL154 as a helper plasmid which contains Komari fragment to facilitate transformation. These five dual binary vector combinations were transformed into Agrobacterium strain AGL1 and used to transform durum wheat cv Stewart 63. Evaluation of the co-transformation efficiencies, the frequencies of marker-free transgenic plants, and integration of backbone sequences in the obtained transgenic lines indicated that two vectors (5G7B and 5TGTB154) were more efficient in generating marker-free transgenic wheat plants with no or minimal integration of backbone sequences in the wheat genome. The vector series developed in this study for generation of marker- and/or backbone-free transgenic wheat plants via Agrobacterium-mediated transformation will be useful to facilitate the creation of “clean” GM wheat containing only the foreign genes of agronomic importance. PMID:27708648

  5. Development of Agrobacterium-mediated transformation of highly valued hill banana cultivar Virupakshi (AAB) for resistance to BBTV disease.

    PubMed

    Elayabalan, Sivalingam; Kalaiponmani, Kalaimughilan; Subramaniam, Sreeramanan; Selvarajan, Ramasamy; Panchanathan, Radha; Muthuvelayoutham, Ramlatha; Kumar, Krish K; Balasubramanian, Ponnuswami

    2013-04-01

    One of the most severe viral diseases of hill banana is caused by banana bunchy top virus (BBTV), a nanovirus transmitted by the aphid Pentalonia nigronervosa. In this study, we reported the Agrobacterium-mediated transformation on a highly valued hill banana cultivar Virupakshi (AAB) for resistance to BBTV disease. The target of the RNA interference (RNAi) is the rep gene, encoded by the BBTV-DNA1. In order to develop RNAi construct targeting the BBTV rep gene, the full-length rep gene of 870 bp was polymerase chain reaction amplified from BBTV infected hill banana sample DNA, cloned and confirmed by DNA sequencing. The partial rep gene fragment was cloned in sense and anti sense orientation in the RNAi intermediate vector, pSTARLING-A. After cloning in pSTARLING-A, the cloned RNAi gene cassette was released by NotI enzyme digestion and cloned into the NotI site of binary vector, pART27. Two different explants, embryogenic cells and embryogenic cell suspension derived microcalli were used for co-cultivation. Selection was done in presence of 100 mg/L kanamycin. In total, 143 putative transgenic hill banana lines were generated and established in green house condition. The presence of the transgenes was confirmed in the selected putative transgenic hill banana lines by PCR and reverse transcription PCR analyses. Transgenic hill banana plants expressing RNAi-BBTV rep were obtained and shown to resist infection by BBTV. The transformed plants are symptomless, and the replication of challenge BBTV almost completely suppressed. Hence, the RNAi mediating resistances were shown to be effective management of BBTV in hill banana.

  6. Parameters influencing Agrobacterium-mediated transformation system in safflower genotypes AKS-207 and PKV Pink.

    PubMed

    Dhumale, Dipti Raghunath; Shingote, Prashant Raghunath; Dudhare, Mahendra Shankarrao; Jadhav, Pravin Vishwanath; Kale, Prashant Bhaskar

    2016-12-01

    Shoot regeneration in safflower (Carthamus tinctorius 'AKS 207' and 'PKV Pink') genetically transformed using Agrobacterium was used for assessing various constraints to the efficiency of transformation including infection period, virulence induction medium, co-cultivation period, bacterial titre, selection regime, and the natural phenolic compound acetosyringone. Transformation frequency was promising with 8-10-day-old cotyledonary leaf explants. Therefore, explants of that age cultured on Agrobacterium minimal medium (AB) containing 100 µM acetosyringone were infected with Agrobacterium (cell titre 0.5 OD600nm) for 15 min followed by 48 h of co-cultivation on kanamycin-enriched medium (50 mg/L). Transformation of the shoots was confirmed using β-glucuronidase (GUS) histochemical assay and polymerase chain reaction (PCR). With the transformation protocol thus optimized, the transformation frequency as determined using GUS assays was 54.0 % for AKS 207 and 47.6 % for PKV Pink. The corresponding figures using PCR were 27.0 and 33.3 %. The transformed shoots required 10-14 weeks of culture initiation but produced very few roots.

  7. Agrobacterium-mediated transformation of rough lemon (Citrus jambhiri Lush) with yeast HAL2 gene

    PubMed Central

    2012-01-01

    Background Rough lemon (Citrus jambhiri Lush.) is the most commonly used Citrus rootstock in south Asia. It is extremely sensitive to salt stress that decreases the growth and yield of Citrus crops in many areas worldwide. Over expression of the yeast halotolerant gene (HAL2) results in increasing the level of salt tolerance in transgenic plants. Results Transformation of rough lemon was carried out by using Agrobacterium tumefaciens strains LBA4404 harboring plasmid pJRM17. Transgenic shoots were selected on kanamycin 100 mg L-1 along with 250 mg L-1 each of cefotaxime and vancomycin for effective inhibition of Agrobacterium growth. The Murashige and Skoog (MS) medium containing 200 μM acetoseryngone (AS) proved to be the best inoculation and co-cultivation medium for transformation. MS medium supplemented with 3 mg L-1 of 6-benzylaminopurine (BA) showed maximum regeneration efficiency of the transformed explants. The final selection of the transformed plants was made on the basis of PCR and Southern blot analysis. Conclusion Rough lemon has been successfully transformed via Agrobacterium tumefaciens with β-glucuronidase (GUS) and HAL2. Various factors affecting gene transformation and regeneration efficiency were also investigated. PMID:22691292

  8. Enhancers of Agrobacterium-mediated Transformation of Tibouchina semidecandra Selected on the Basis of GFP Expression.

    PubMed

    Yong, Wilson Thau Lym; Henry, Erle Stanley; Abdullah, Janna Ong

    2010-12-01

    Genetic engineering is a powerful tool for the improvement of plant traits. Despite reported successes in the plant kingdom, this technology has barely scratched the surface of the Melastomataceae family. Limited studies have led to some optimisation of parameters known to affect the transformation efficiency of these plants. The major finding of this study was to optimise the presence of selected enhancers [e.g., monosaccharides (D-glucose, D-galactose and D-fructose), tyrosine, aluminium chloride (AICI3) and ascorbic acid] to improve the transformation efficiency of Tibouchina semidecandra. Agrobacterium tumefaciens strain LBA4404 harbouring the disarmed plasmid pCAMBIA1304 was used to transform shoots and nodes of T. semidecandra. Different concentrations of the transformation enhancers were tested by using green fluorescent protein (GFP) as a reporter. The results obtained were based on the percentage of GFP expression, which was observed 14 days post-transformation. A combination of 120 μM galactose and 100 μM tyrosine supplemented with 600 μM AICI3 in the presence of 15 mg/l ascorbic acid gave the highest percentage of positive transformants for T. semidecandra shoots. Whereas 60 μM galactose and 50 μM tyrosine with 200 μM AICI3 in the presence of 15 mg/l ascorbic acid was optimum for T. semidecandra nodes. The presence of the hygromycin phosphotransferase II (hptII) transgene in the genomic DNA of putative T. semidecandra transformants was verified by PCR amplification with specific primers.

  9. Transgenic grasspea (Lathyrus sativus L.): factors influencing agrobacterium-mediated transformation and regeneration.

    PubMed

    Barik, D P; Mohapatra, U; Chand, P K

    2005-11-01

    A reproducible procedure was developed for genetic transformation of grasspea using epicotyl segment co-cultivation with Agrobacterium. Two disarmed Agrobacterium tumefaciens strains, EHA 105 and LBA 4404, both carrying the binary plasmid p35SGUSINT with the neomycin phosphotransferase II (nptII) gene and the beta-glucuronidase (gus)-intron, were studied as vector systems. The latter was found to have a higher transforming ability. Several key factors modifying the transformation rate were optimized. The highest transformation rate was achieved using hand-pricked explants for infection with an Agrobacterium culture corresponding to OD(600) congruent with 0.6 and diluted to a cell density of 10(9) cells ml(-1) for 10 min, followed by co-cultivation for 4 days in a medium maintained at pH 5.6. Putative transformed explants capable of forming shoots were selected on regeneration medium containing kanamycin (100 mug ml(-1)). We achieved up to 36% transient expression based on the GUS histochemical assay. Southern hybridization of genomic DNA of the kanamycin-resistant GUS-expressive shoots to a gus-intron probe substantiated the integration of the transgene. Transformed shoots were rooted on half-strength MS containing 0.5 mg l(-1) indole-3-acetic acid, acclimated in vermi-compost and established in the experimental field. Germ-line transformation was evident through progeny analysis. Among T(1) seedlings of most transgenic plant lines, kanamycin-resistant and -sensitive plants segregated in a ratio close to 3:1.

  10. Agrobacterium-mediated transformation of promising oil-bearing marine algae Parachlorella kessleri.

    PubMed

    Rathod, Jayant Pralhad; Prakash, Gunjan; Pandit, Reena; Lali, Arvind M

    2013-11-01

    Parachlorella kessleri is a unicellular alga which grows in fresh as well as marine water and is commercially important as biomass/lipid feedstock and in bioremediation. The present study describes the successful transformation of marine P. kessleri with the help of Agrobacterium tumefaciens. Transformed marine P. kessleri was able to tolerate more than 10 mg l(-1) hygromycin concentration. Co-cultivation conditions were modulated to allow the simultaneous growth of both marine P. kessleri and A. tumefaciens. For co-cultivation, P. kessleri was shifted from Walne's to tris acetate phosphate medium to reduce the antibiotic requirement during selection. In the present study, the transfer of T-DNA was successful without using acetosyringone. Biochemical and genetic analyses were performed for expression of transgenes by GUS assay and PCR in transformants. Establishment of this protocol would be useful in further genetic modification of oil-bearing Parachlorella species.

  11. Agrobacterium-mediated transformation of Easter lily (Lilium longiflorum cv. Nellie White)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Conditions were optimized for transient transformation of Lilium longiflorum cv. Nellie White using Agrobacterium tumefaciens. Bulb scale and basal meristem explants were inoculated with A. tumefaciens strain AGL1 containing the binary vector pCAMBIA 2301 which has the uidA gene that codes for ß-gl...

  12. Agrobacterium-mediated transformation of polyploid cereals. The efficiency of selection and transgene expression in wheat.

    PubMed

    Przetakiewicz, Anna; Karaś, Agnieszka; Orczyk, Wacław; Nadolska-Orczyk, Anna

    2004-01-01

    Three combinations of Agrobacterium tumefaciens strains and vectors were used in the transformation of selected Polish wheat cultivars. The combinations were: two hypervirulent strains, AGL1, containing the pDM805 binary plasmid, and EHA101, containing pGAH; and the common Agro strain LBA4404, harboring the super-binary pTOK233 vector. pDM805 contained bar under the control of Ubi1 promoter, pGAH had nptII under nos, and pTOK233 had hpt under 35S. Additionally, pDM805 and pTOK233 carried the gus reporter gene under the Act1 promoter or 35S promoter, respectively. The highest selection rate was 12.6% and was obtained with EHA101(pGAH) on a kanamycin-containing medium. Sixty-five of the plants grown on that medium were PCR positive. The second best combination was LBA4404(pTOK233) and kanamycin selection, which gave an average transformation rate of 2.3%. Phosphinothricin selection gave 1.0% transformation efficiency, while hygromycin, depending on the strain/vector used, gave from 0.2 to 0.4%. PCR tests in T1 revealed that 67% of the lines showed a 3:1 segregation ratio, and 11% a 15:1 ratio, while in 22%, segregation was non-Mendelian. The high number of T0 transgenic plants containing one copy of the transgene was confirmed via Southern blot analysis. Kanamycin resistance in the T1 generation was very low; in some lines, all the progeny were kanamycin sensitive. GUS expression, only tested in young T1 plants, was in agreement with Mendelian segregation in three out of the twelve tested. The factors influencing the efficiency of selection and transgene expression are discussed in this paper.

  13. An efficient plant regeneration and Agrobacterium-mediated genetic transformation of Tagetes erecta.

    PubMed

    Gupta, Vijayta; Ur Rahman, Laiq

    2015-07-01

    Tagetes erecta, L. an asteraceous plant of industrial and medicinal value, contains important compounds like pyrethrins, thiophenes and lutein, possessing immense potential for insecticidal, nematicidal and nutraceutical activities. Considering the importance and demand for these natural compounds, genetic manipulation of this crop for better productivity of secondary metabolites holds great significance. A rapid and reproducible direct regeneration and genetic transformation system is the prerequisite for genetic manipulation of any crop. This paper elucidates the establishment of an efficient direct regeneration and transformation protocol of T. erecta using Agrobacterium tumefaciens. Investigation of the effects of different types of explants (Hypocotyls, cotyledonary leaves, rachis and leaf sections) and different BAP and GA3 combinations on the regeneration frequency of T. erecta suggested that the best regeneration frequency (66 %) with an average of 5.08 ± 0.09 shoot buds/explant was observed from hypocotyl explants cultured on media containing 1.5 mg/l BAP and 5 mg/l GA3. The transformation protocol was established using A. tumefaciens strain LBA4404, containing the binary vector pBI121, along with the gusA reporter gene with intron under the transcriptional control of the Cauliflower Mosaic Virus (CaMV) 35S promoter and the neomycin phosphotransferase II (nptII) gene as a kanamycin-resistant plant-selectable marker. Various parameters like optimization of kanamycin concentration (200 mg/l) for selection, standardization of cocultivation time (45 min) and acetosyringone concentration (150 μM) for obtaining higher transformation frequency were established using hypocotyl explants. The selected putative transgenic shoots were subsequently rooted on the Murashige and Skoog medium and transferred to the green house successfully. The plants were characterised by analysing the gus expression, amplification of 600 bp npt II fragment and Southern blot

  14. Generation of marker-free transgenic hexaploid wheat via an Agrobacterium-mediated co-transformation strategy in commercial Chinese wheat varieties.

    PubMed

    Wang, Ke; Liu, Huiyun; Du, Lipu; Ye, Xingguo

    2016-11-10

    Genotype specificity is a big problem lagging the development of efficient hexaploid wheat transformation system. Increasingly, the biosecurity of genetically modified organisms is garnering public attention, so the generation of marker-free transgenic plants is very important to the eventual potential commercial release of transgenic wheat. In this study, 15 commercial Chinese hexaploid wheat varieties were successfully transformed via an Agrobacterium-mediated method, with efficiency of up to 37.7%, as confirmed by the use of Quickstix strips, histochemical staining, PCR analysis and Southern blotting. Of particular interest, marker-free transgenic wheat plants from various commercial Chinese varieties and their F1 hybrids were successfully obtained for the first time, with a frequency of 4.3%, using a plasmid harbouring two independent T-DNA regions. The average co-integration frequency of the gus and the bar genes located on the two independent T-DNA regions was 49.0% in T0 plants. We further found that the efficiency of generating marker-free plants was related to the number of bar gene copies integrated in the genome. Marker-free transgenic wheat plants were identified in the progeny of three transgenic lines that had only one or two bar gene copies. Moreover, silencing of the bar gene was detected in 30.7% of T1 positive plants, but the gus gene was never found to be silenced in T1 plants. Bisulphite genomic sequencing suggested that DNA methylation in the 35S promoter of the bar gene regulatory region might be the main reason for bar gene silencing in the transgenic plants.

  15. A new high-frequency Agrobacterium-mediated transformation technique for Sesamum indicum L. using de-embryonated cotyledon as explant.

    PubMed

    Chowdhury, Supriyo; Basu, Arpita; Kundu, Surekha

    2014-09-01

    In spite of the economic importance of sesame (Sesamum indicum L.) and the recent availability of its genome sequence, a high-frequency transformation protocol is still not available. The only two existing Agrobacterium-mediated transformation protocols that are available have poor transformation efficiencies of less than 2%. In the present study, we report a high-frequency, simple, and reproducible transformation protocol for sesame. Transformation was done using de-embryonated cotyledons via somatic embryogenic stages. All the critical parameters of transformation, like incubation period of explants in pre-regeneration medium prior to infection by Agrobacterium tumefaciens, cocultivation period, concentrations of acetosyringone in cocultivation medium, kanamycin concentration, and concentration of plant hormones, including 6-benzylaminopurine, have been optimized. This protocol is superior to the two existing protocols in its high regeneration and transformation efficiencies. The transformed sesame lines have been tested by PCR, RT-PCR for neomycin phosphotransferase II gene expression, and β-glucuronidase (GUS) assay. The regeneration frequency and transformation efficiency are 57.33 and 42.66%, respectively. T0 and T1 generation transgenic plants were analyzed, and several T1 plants homozygous for the transgenes were obtained.

  16. A comparison of Agrobacterium-mediated transformation and protoplast-mediated transformation with CRISPR-Cas9 and bipartite gene targeting substrates, as effective gene targeting tools for Aspergillus carbonarius.

    PubMed

    Weyda, István; Yang, Lei; Vang, Jesper; Ahring, Birgitte K; Lübeck, Mette; Lübeck, Peter S

    2017-04-01

    In recent years, versatile genetic tools have been developed and applied to a number of filamentous fungi of industrial importance. However, the existing techniques have limitations when it comes to achieve the desired genetic modifications, especially for efficient gene targeting. In this study, we used Aspergillus carbonarius as a host strain due to its potential as a cell factory, and compared three gene targeting techniques by disrupting the ayg1 gene involved in the biosynthesis of conidial pigment in A. carbonarius. The absence of the ayg1 gene leads to phenotypic change in conidia color, which facilitated the analysis on the gene targeting frequency. The examined transformation techniques included Agrobacterium-mediated transformation (AMT) and protoplast-mediated transformation (PMT). Furthermore, the PMT for the disruption of the ayg1 gene was carried out with bipartite gene targeting fragments and the recently adapted CRISPR-Cas9 system. All three techniques were successful in generating Δayg1 mutants, but showed different efficiencies. The most efficient method for gene targeting was AMT, but further it was shown to be dependent on the choice of Agrobacterium strain. However, there are different advantages and disadvantages of all three gene targeting methods which are discussed, in order to facilitate future approaches for fungal strain improvements.

  17. SacB-SacR Gene Cassette As the Negative Selection Marker to Suppress Agrobacterium Overgrowth in Agrobacterium-Mediated Plant Transformation

    PubMed Central

    Liu, Yiming; Miao, Jiamin; Traore, Sy; Kong, Danyu; Liu, Yi; Zhang, Xunzhong; Nimchuk, Zachary L.; Liu, Zongrang; Zhao, Bingyu

    2016-01-01

    Agrobacterium overgrowth is a common problem in Agrobacterium-mediated plant transformation. To suppress the Agrobacterium overgrowth, various antibiotics have been used during plant tissue culture steps. The antibiotics are expensive and may adversely affect plant cell differentiation and reduce plant transformation efficiency. The SacB-SacR proteins are toxic to most Agrobacterium tumefaciens strains when they are grown on culture medium supplemented with sucrose. Therefore, SacB-SacR genes can be used as negative selection markers to suppress the overgrowth of A. tumefaciens in the plant tissue culture process. We generated a mutant A. tumefaciens strain GV2260 (recA-SacB/R) that has the SacB-SacR cassette inserted into the bacterial genome at the recA gene locus. The mutant Agrobacterium strain is sensitive to sucrose but maintains its ability to transform plant cells in both transient and stable transformation assays. We demonstrated that the mutant strain GV2260 (recA-SacB/R) can be inhibited by sucrose that reduces the overgrowth of Agrobacterium and therefore improves the plant transformation efficiency. We employed GV2260 (recA-SacB/R) to generate stable transgenic N. benthamiana plants expressing a CRISPR-Cas9 for knocking out a WRKY transcription factor. PMID:27833912

  18. SacB-SacR Gene Cassette As the Negative Selection Marker to Suppress Agrobacterium Overgrowth in Agrobacterium-Mediated Plant Transformation.

    PubMed

    Liu, Yiming; Miao, Jiamin; Traore, Sy; Kong, Danyu; Liu, Yi; Zhang, Xunzhong; Nimchuk, Zachary L; Liu, Zongrang; Zhao, Bingyu

    2016-01-01

    Agrobacterium overgrowth is a common problem in Agrobacterium-mediated plant transformation. To suppress the Agrobacterium overgrowth, various antibiotics have been used during plant tissue culture steps. The antibiotics are expensive and may adversely affect plant cell differentiation and reduce plant transformation efficiency. The SacB-SacR proteins are toxic to most Agrobacterium tumefaciens strains when they are grown on culture medium supplemented with sucrose. Therefore, SacB-SacR genes can be used as negative selection markers to suppress the overgrowth of A. tumefaciens in the plant tissue culture process. We generated a mutant A. tumefaciens strain GV2260 (recA-SacB/R) that has the SacB-SacR cassette inserted into the bacterial genome at the recA gene locus. The mutant Agrobacterium strain is sensitive to sucrose but maintains its ability to transform plant cells in both transient and stable transformation assays. We demonstrated that the mutant strain GV2260 (recA-SacB/R) can be inhibited by sucrose that reduces the overgrowth of Agrobacterium and therefore improves the plant transformation efficiency. We employed GV2260 (recA-SacB/R) to generate stable transgenic N. benthamiana plants expressing a CRISPR-Cas9 for knocking out a WRKY transcription factor.

  19. Development of an efficient Agrobacterium-mediated transformation system and production of herbicide-resistant transgenic plants in garlic (Allium sativum L.).

    PubMed

    Ahn, Yul-Kyun; Yoon, Moo-Kyoung; Jeon, Jong-Seong

    2013-08-01

    The genetic improvement of garlic plants (Allium sativum L.) with agronomical beneficial traits is rarely achieved due to the lack of an applicable transformation system. Here, we developed an efficient Agrobacterium-mediated transformation procedure with Danyang, an elite Korean garlic cultivar. Examination of sGFP (synthetic green fluorescence protein) expression revealed that treatment with 2-(N-morpholino) ethanesulfonic acid (MES), L-cysteine and/or dithiothreitol (DTT) gives the highest efficiency in transient gene transfer during Agrobacterium co-cultivation with calli derived from the roots of in vitro plantlets. To increase stable transformation efficiency, a two-step selection was employed on the basis of hygromycin resistance and sGFP expression. Of the hygromycin-resistant calli initially produced, only sGFP-expressing calli were subcultured for selection of transgenic calli. Transgenic plantlets produced from these calli were grown to maturity. The transformation efficiency increased up to 10.6% via our optimized procedure. DNA and RNA gel-blot analysis indicated that transgenic garlic plants stably integrated and expressed the phosphinothricin acetyltransferase (PAT) gene. A herbicide spraying assay demonstrated that transgenic plants of garlic conferred herbicide resistance, whilst nontransgenic plants and weeds died. These results indicate that our transformation system can be efficiently utilized to produce transgenic garlic plants with agronomic benefits.

  20. Agrobacterium-mediated plant transformation by novel mini-T vectors in conjunction with a high-copy vir region helper plasmid.

    PubMed

    Zyprian, E; Kado, C I

    1990-08-01

    A new binary vector system for Agrobacterium-mediated plant transformation was developed. A set of four mini-T vectors comprised of T-DNA border sequences from nopaline-type Ti-plasmid pTiC58 flanking a chimaeric hygromycin-resistance gene for selection of transformants and up to eight unique restriction sites for cloning foreign DNA was constructed on a broad-host replicon containing the oriV of plasmid pSa. In two of the constructs these multiple cloning sites are flanked by a strong promoter to activate transcription of inserted DNA in planta. High-efficiency transformation was prompted by a high-copy, stable virulence helper plasmid pUCD2614, which contains a cloned virulence region of pTiC58 and tandem copies of the par locus of plasmid pTAR. Southern blot hybridization and genetic analyses of the progeny of transformed plants showed that the hygromycin resistance gene was stably inherited.

  1. Assessment of factors influencing the Agrobacterium-mediated in planta seed transformation of brinjal (Solanum melongena L.).

    PubMed

    Subramanyam, Kondeti; Rajesh, Manoharan; Jaganath, Balusamy; Vasuki, Amirthalingam; Theboral, Jeevaraj; Elayaraja, Dhandapani; Karthik, Sivabalan; Manickavasagam, Markandan; Ganapathi, Andy

    2013-09-01

    An efficient and reproducible in planta transformation method was developed for brinjal using seed as an explant. The brinjal seeds were infected with Agrobacterium tumefaciens EHA 105 harbouring pCAMBIA 1301-bar plasmid, and the transformants were selected against BASTA®. Several parameters influencing the in planta seed transformation such as pre-culture duration, acetosyringone concentration, surfactants, duration of sonication, vacuum pressure and vacuum duration have been evaluated. The putatively transformed (T 0) brinjal plants were screened by GUS histochemical analysis. Among the different combinations and concentrations tested, when the 18-h pre-cultured brinjal seeds were sonicated for 20 min and vacuum infiltered for 3 min at 500 mm of Hg in Agrobacterium suspension containing 100 μM acetosyringone, 0.2 % Silwett L-77 favoured the Agrobacterium infection and showed maximum transformation efficiency. Among the five brinjal varieties evaluated, Arka Samhitha showed maximum transformation efficiency at 45.66 %. The transgene was successfully transmitted to progeny plants (T 1) which was evidenced by GUS histochemical analysis, polymerase chain reaction and Southern hybridisation. The in planta protocol developed in the present study would be beneficial to transfer the economically and nutritionally important genes into different varieties of brinjal, and the transgenic brinjal plants can be produced in less time (approximately 27 days).

  2. The Agrobacterium-mediated transformation of common wheat (Triticum aestivum L.) and triticale (x Triticosecale Wittmack): role of the binary vector system and selection cassettes.

    PubMed

    Bińka, Agnieszka; Orczyk, Wacław; Nadolska-Orczyk, Anna

    2012-02-01

    The influence of two binary vector systems, pGreen and pCAMBIA, on the Agrobacterium-mediated transformation ability of wheat and triticale was studied. Both vectors carried selection cassettes with bar or nptII driven by different promoters. Two cultivars of wheat, Kontesa and Torka, and one cultivar of triticale, Wanad, were tested. The transformation rates for the wheat cultivars ranged from 0.00 to 3.58% and from 0.00 to 6.79% for triticale. The best values for wheat were 3.58% for Kontesa and 3.14% for Torka, and these were obtained after transformation with the pGreen vector carrying the nptII selection gene under the control of 35S promoter. In the case of the bar selection system, the best transformation rates were, respectively, 1.46 and 1.79%. Such rates were obtained when the 35S::bar cassette was carried by the pCAMBIA vector; they were significantly lower with the pGreen vector. The triticale cultivar Wanad had its highest transformation rate after transformation with nptII driven by 35S in pCAMBIA. The bar selection system for the same triticale cultivar was better when the gene was driven by nos and the selection cassette was carried by pGreen. The integration of the transgenes was confirmed with at least three pairs of specific starters amplifying the fragments of nptII, bar, or gus. The expression of selection genes, measured by reverse transcriptase polymerase chain reaction (RT-PCR) in relation to the actin gene, was low, ranging from 0.00 to 0.63 for nptII and from 0.00 to 0.33 for bar. The highest relative transcript accumulation was observed for nptII driven by 35S and expressed in Kontesa that had been transformed with pGreen.

  3. A simple shoot multiplication procedure using internode explants, and its application for particle bombardment and Agrobacterium-mediated transformation in watercress.

    PubMed

    Ogita, Shinjiro; Usui, Miki; Shibutani, Nanae; Kato, Yasuo

    2009-07-01

    A shoot multiplication system derived from internode explants was investigated with the aim of improving genetic characteristics of watercress (Nasturtium officinale R. Br.). Internodes of ca. 1 cm excised from in vitro stock shoot culture were placed on half-strength Murashige and Skoog (MS) medium supplemented with 3 muM 2,4-dichlorophenoxyacetic acid as a pre-treatment. Laser scanning microscopy indicated clearly that the first sign of meristematic cell division could be seen after 1-2 days of pre-culture, and meristematic tissues multiplied along the vascular cambium of the internode segment during 7 days of culture. Multiple shoots could be obtained from more than 90% of the pre-treated explants when they were subsequently transferred to MS medium supplemented with 1 muM thidiazuron for 3 weeks. These findings indicate that pre-treatment of the internodes for 7 days promoted their capacity for organogenesis. Using this pre-treatment, frequent generation of transgenic watercress plants was achieved by adapting particle bombardment and Agrobacterium-mediated transformation techniques with a construct expressing a synthetic green florescent protein gene.

  4. Highly efficient Agrobacterium-mediated transformation of embryogenic cell suspensions of Musa acuminata cv. Mas (AA) via a liquid co-cultivation system.

    PubMed

    Huang, Xia; Huang, Xue-Lin; Xiao, Wang; Zhao, Jie-Tang; Dai, Xue-Mei; Chen, Yun-Feng; Li, Xiao-Ju

    2007-10-01

    A high efficient protocol of Agrobacterium-mediated transformation of Musa acuminata cv. Mas (AA), a major banana variety of the South East Asia region, was developed in this study. Male-flower-derived embryogenic cell suspensions (ECS) were co-cultivated in liquid medium with Agrobacterium strain EHA105 harboring a binary vector pCAMBIA2301 carrying nptII and gusA gene in the T-DNA. Depending upon conditions and duration of co-cultivation in liquid medium, 0-490 transgenic plants per 0.5 ml packed cell volume (PCV) of ECS were obtained. The optimum duration of inoculation was 2 h, and the highest transformation frequency was achieved when infected ECS were co-cultivated in liquid medium first for 12 h at 40 rpm and then for 156 h at 100 rpm on a rotary shaker. Co-cultivation for a shorter duration (72 h) or shaking constantly at 100 rpm at the same duration gave 1.6 and 1.8 folds lower transformation efficiency, respectively. No transgenic plants were obtained in parallel experiments carried on semi-solid media. Histochemical GUS assay and molecular analysis in several tissues of the transgenic plants demonstrated that foreign genes were stably integrated into the banana genome. Compared to semi-solid co-cultivation transformation in other banana species, it is remarkable that liquid co-cultivation was much more efficient for transformation of the Mas cultivar, and was at least 1 month faster for regenerating transgenic plants.

  5. Improvement in Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) by the inhibition of polyphenolics released during wounding of cotyledonary node explants.

    PubMed

    Yadav, Reena; Mehrotra, Meenakshi; Singh, Aditya K; Niranjan, Abhishek; Singh, Rani; Sanyal, Indraneel; Lehri, Alok; Pande, Veena; Amla, D V

    2017-01-01

    Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) has been performed using cotyledonary node explants (CNs), which release phenolics upon excision that are detrimental to the viability of Agrobacterium tumefaciens and result in low transformation frequency. Twelve low molecular weight phenolic compounds and salicylic acid were identified in the exudates released upon excision during the preparation of cotyledonary nodes by reverse phase high-performance liquid chromatography (RP-HPLC). Zone inhibition assays performed with the explant exudates released at periodic intervals after excision showed the inhibition of A. tumefaciens. Agroinoculation of freshly excised cotyledonary nodes of chickpea showed 98-99 % inhibition of colony forming units (cfu). Osmium tetraoxide fixation of excised tissues showed enhanced accumulation of phenolics in the sub-epidermal regions causing enzymatic browning, affecting the viability and performance of A. tumefaciens for T-DNA delivery. The periodic analysis of exudates released from excised CNs showed enhanced levels of gallic acid (0.2945 ± 0.014 μg/g), chlorogenic acid (0.0978 ± 0.0046 μg/g), and quercetin (0.0971 ± 0.0046 μg/g) fresh weight, which were detrimental to A. tumefaciens. Quantitative assays and the elution profile showed the maximum leaching of phenolics, flavonoids, and salicylic acid immediately after the excision of explants and continued till 4 to 8 h post-excision. Pre-treatment of excised explants with inhibitors of polyphenol oxidase like L-cysteine, DTT, and sodium thiosulfate before co-cultivation showed the recovery of A. tumefaciens cfu, decreased the accumulation of phenolics, and improved transformation frequency. Our results show the hypersensitive response of excision stress for the expression of defense response-related genes and synthesis of metabolites in grain legume chickpea against pathogen infestation including Agrobacterium.

  6. Agrobacterium-mediated transformation of cotton (Gossypium hirsutum) shoot apex with a fungal phytase gene improves phosphorus acquisition.

    PubMed

    Ma, Zhiying; Liu, Jianfeng; Wang, Xingfen

    2013-01-01

    Cotton is an important world economic crop plant. It is considered that cotton is recalcitrant to in vitro proliferation. Somatic embryogenesis and plant regeneration has been successful by using hypocotyl, whereas it is highly genotype dependent. Here, a genotype-independent cotton regeneration protocol from shoot apices is presented. Shoot apices from 3- to 5-day-old seedlings of cotton are infected with an Agrobacterium strain, EHA105, carrying the binary vector pC-KSA contained phytase gene (phyA) and the marker gene neomycin phosphotransferase (NPTII), and directly regenerated as shoots in vitro. Rooted shoots can be obtained within 6-8 weeks. Plants that survived by leaf painting kanamycin (kan) were -further analyzed by DNA and RNA blottings. The transgenic plants with increased the phosphorus (P) acquisition efficiency were obtained following the transformation method.

  7. Development of an Agrobacterium-mediated transformation protocol for the tree-legume Leucaena leucocephala using immature zygotic embryos.

    PubMed

    Jube, Sandro; Borthakur, Dulal

    2009-01-01

    The tree-legume Leucaena leucocephala (leucaena) is used as a perennial fodder because of its fast-growing foliage, which is high in protein content. The use of leucaena as a fodder is however restricted due to the presence of the toxin mimosine. Improvements in the nutritional contents as well as other agronomic traits of leucaena can be accomplished through genetic transformation. The objective of this research was to develop a transformation protocol for leucaena using phosphinothricin resistance as the plant selectable marker. Explants obtained from immature zygotic embryos infected with the Agrobacterium tumefaciens strain C58C1 containing the binary plasmid pCAMBIA3201 produced four putative transformed leucaena plants. Transformation was con- firmed by PCR, RT-PCR, Southern blot, Western analyses, GUS-specific enzyme activity and herbicide leaf spraying assay. A transformation efficiency of 2% was established using this protocol.

  8. High-efficiency Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) and regeneration of insect-resistant transgenic plants.

    PubMed

    Mehrotra, Meenakshi; Sanyal, Indraneel; Amla, D V

    2011-09-01

    To develop an efficient genetic transformation system of chickpea (Cicer arietinum L.), callus derived from mature embryonic axes of variety P-362 was transformed with Agrobacterium tumefaciens strain LBA4404 harboring p35SGUS-INT plasmid containing the uidA gene encoding β-glucuronidase (GUS) and the nptII gene for kanamycin selection. Various factors affecting transformation efficiency were optimized; as Agrobacterium suspension at OD(600) 0.3 with 48 h of co-cultivation period at 20°C was found optimal for transforming 10-day-old MEA-derived callus. Inclusion of 200 μM acetosyringone, sonication for 4 s with vacuum infiltration for 6 min improved the number of GUS foci per responding explant from 1.0 to 38.6, as determined by histochemical GUS assay. For introducing the insect-resistant trait into chickpea, binary vector pRD400-cry1Ac was also transformed under optimized conditions and 18 T(0) transgenic plants were generated, representing 3.6% transformation frequency. T(0) transgenic plants reflected Mendelian inheritance pattern of transgene segregation in T(1) progeny. PCR, RT-PCR, and Southern hybridization analysis of T(0) and T(1) transgenic plants confirmed stable integration of transgenes into the chickpea genome. The expression level of Bt-Cry protein in T(0) and T(1) transgenic chickpea plants was achieved maximum up to 116 ng mg(-1) of soluble protein, which efficiently causes 100% mortality to second instar larvae of Helicoverpa armigera as analyzed by an insect mortality bioassay. Our results demonstrate an efficient and rapid transformation system of chickpea for producing non-chimeric transgenic plants with high frequency. These findings will certainly accelerate the development of chickpea plants with novel traits.

  9. Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14.

    PubMed

    Nyaboga, Evans N; Njiru, Joshua M; Tripathi, Leena

    2015-01-01

    Routine production of large numbers of transgenic plants is required to fully exploit advances in cassava biotechnology and support development of improved germplasm for deployment to farmers. This article describes an improved, high-efficiency transformation protocol for recalcitrant cassava cultivar TME14 preferred in Africa. Factors that favor production of friable embryogenic calli (FEC) were found to be use of DKW medium, crushing of organized embryogenic structures (OES) through 1-2 mm sized metal wire mesh, washing of crushed OES tissues and short exposure of tyrosine to somatic embryos; and transformation efficiency was enhanced by use of low Agrobacterium density during co-cultivation, co-centrifugation of FEC with Agrobacterium, germination of paramomycin resistant somatic embryos on medium containing BAP with gradual increase in concentration and variations of the frequency of subculture of cotyledonary-stage embryos on shoot elongation medium. By applying the optimized parameters, FEC were produced for cassava cultivar TME14 and transformed using Agrobacterium strain LBA4404 harboring the binary vector pCAMBIA2301. About 70-80 independent transgenic lines per ml settled cell volume (SCV) of FEC were regenerated on selective medium. Histochemical GUS assays confirmed the expression of gusA gene in transformed calli, somatic embryos and transgenic plants. The presence and integration of the gusA gene were confirmed by PCR and Southern blot analysis, respectively. RT-PCR analysis of transgenic plants confirmed the expression of gusA gene. This protocol demonstrates significantly enhanced transformation efficiency over existing cassava transformation protocols and could become a powerful tool for functional genomics and transferring new traits into cassava.

  10. Factors influencing somatic embryogenesis, regeneration, and Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) cultivar TME14

    PubMed Central

    Nyaboga, Evans N.; Njiru, Joshua M.; Tripathi, Leena

    2015-01-01

    Routine production of large numbers of transgenic plants is required to fully exploit advances in cassava biotechnology and support development of improved germplasm for deployment to farmers. This article describes an improved, high-efficiency transformation protocol for recalcitrant cassava cultivar TME14 preferred in Africa. Factors that favor production of friable embryogenic calli (FEC) were found to be use of DKW medium, crushing of organized embryogenic structures (OES) through 1–2 mm sized metal wire mesh, washing of crushed OES tissues and short exposure of tyrosine to somatic embryos; and transformation efficiency was enhanced by use of low Agrobacterium density during co-cultivation, co-centrifugation of FEC with Agrobacterium, germination of paramomycin resistant somatic embryos on medium containing BAP with gradual increase in concentration and variations of the frequency of subculture of cotyledonary-stage embryos on shoot elongation medium. By applying the optimized parameters, FEC were produced for cassava cultivar TME14 and transformed using Agrobacterium strain LBA4404 harboring the binary vector pCAMBIA2301. About 70–80 independent transgenic lines per ml settled cell volume (SCV) of FEC were regenerated on selective medium. Histochemical GUS assays confirmed the expression of gusA gene in transformed calli, somatic embryos and transgenic plants. The presence and integration of the gusA gene were confirmed by PCR and Southern blot analysis, respectively. RT-PCR analysis of transgenic plants confirmed the expression of gusA gene. This protocol demonstrates significantly enhanced transformation efficiency over existing cassava transformation protocols and could become a powerful tool for functional genomics and transferring new traits into cassava. PMID:26113851

  11. [Effective agrobacterium-mediated transformation of chicory (Cichorium intybus L.) using vector with gene of tuberculosis antigene ESAT6].

    PubMed

    Matveeva, N A; Vasilenko, M Iu; Shakhovskiĭ, A M; Bannikova, M A; Kvasko, O Iu; Kuchuk, N V

    2011-01-01

    The conditions of high efficient chicory transformation with Mycobacterium tuberculosis antigene ESAT6 have been determined. Transformation frequency was up to 86% when the cotyledons were cultivated within 3 days without cefotaxime and then 1 day without kanamycine. DNA PCR-analysis has shown the presence both of selective nptII and target esxA genes in all analysed plants. At the same time RT-PCR has shown the presence of nptII transcripts for eight analysed lines and esxA transcripts for only five analysed lines.

  12. Sonication, Vacuum Infiltration and Thiol Compounds Enhance the Agrobacterium-Mediated Transformation Frequency of Withania somnifera (L.) Dunal

    PubMed Central

    Sivanandhan, Ganeshan; Kapil Dev, Gnajothi; Theboral, Jeevaraj; Selvaraj, Natesan; Ganapathi, Andy; Manickavasagam, Markandan

    2015-01-01

    In the present study, we have established a stable transformation protocol via Agrobacterium tumafacines for the pharmaceutically important Withania somnifera. Six day-old nodal explants were used for 3 day co-cultivation with Agrobacterium tumefaciens strain LBA4404 harbouring the vector pCAMIBA2301. Among the different injury treatments, sonication, vacuum infiltration and their combination treatments tested, a vacuum infiltration for 10 min followed by sonication for 10 sec with A. tumefaciens led to a higher transient GUS expression (84% explants expressing GUS at regenerating sites). In order to improve gene integration, thiol compounds were added to co-cultivation medium. A combined treatment of L-Cys at 100 mg/l, STS at 125 mg/l, DTT at 75 mg/l resulted in a higher GUS expression (90%) in the nodal explants. After 3 days of co-cultivation, the explants were subjected to three selection cycles with increasing concentrations of kanamycin [100 to 115 mg/l]. The integration and expression of gusA gene in T0 and T1 transgenic plants were confirmed by polymerase chain reaction (PCR), and Southern blott analysis. These transformed plants (T0 and T1) were fertile and morphologically normal. From the present investigation, we have achieved a higher transformation efficiency of (10%). Withanolides (withanolide A, withanolide B, withanone and withaferin A) contents of transformed plants (T0 and T1) were marginally higher than control plants. PMID:25927703

  13. The usefulness of the gfp reporter gene for monitoring Agrobacterium-mediated transformation of potato dihaploid and tetraploid genotypes.

    PubMed

    Rakosy-Tican, Elena; Aurori, Cristian M; Dijkstra, Camelia; Thieme, Ramona; Aurori, Adriana; Davey, Michael R

    2007-05-01

    Potato is one of the main targets for genetic improvement by gene transfer. The aim of the present study was to establish a robust protocol for the genetic transformation of three dihaploid and four economically important cultivars of potato using Agrobacterium tumefaciens carrying the in vivo screenable reporter gene for green fluorescent protein (gfp) and the marker gene for neomycin phosphotransferase (nptII). Stem and leaf explants were used for transformation by Agrobacterium tumefaciens strain LBA4404 carrying the binary vector pHB2892. Kanamycin selection, visual screening of GFP by epifluorescent microscopy, PCR amplification of nptII and gfp genes, as well as RT-PCR and Southern blotting of gfp and Northern blotting of nptII, were used for transgenic plant selection, identification and analysis. Genetic transformation was optimized for the best performing genotypes with a mean number of shoots expressing gfp per explant of 13 and 2 (dihaploid line 178/10 and cv. 'Baltica', respectively). The nptII marker and gfp reporter genes permitted selection and excellent visual screening of transgenic tissues and plants. They also revealed the effects of antibiotic selection on organogenesis and transformation frequency, and the identification of escapes and chimeras in all potato genotypes. Silencing of the gfp transgene that may represent site-specific inactivation during cell differentiation, occurred in some transgenic shoots of tetraploid cultivars and in specific chimeric clones of the dihaploid line 178/10. The regeneration of escapes could be attributed to either the protection of non-transformed cells by neighbouring transgenic cells, or the persistence of Agrobacterium cells in plant tissues after co-cultivation.

  14. Transgenic superroots of Lotus corniculatus can be regenerated from superroot-derived leaves following Agrobacterium-mediated transformation.

    PubMed

    Tanaka, Hidenori; Toyama, Jun; Hashiguchi, Masatsugu; Kutsuna, Yasuyo; Tsuruta, Shin-ichi; Akashi, Ryo; Hoffmann, Franz

    2008-08-25

    Super-growing roots (superroots; SR), which have been established in the legume species Lotus corniculatus, are a fast-growing root culture that allows continuous root cloning, direct somatic embryogenesis and mass regeneration of plants under entirely growth regulator-free culture conditions. These features are unique for non-hairy root cultures, and they are now stably expressed since the culture was isolated more than 10 years ago (1997). Attempts to achieve direct and stable transformation of SR turned out to be unsuccessful. Making use of the supple regeneration plasticity of SR, we are reporting here an indirect transformation protocol. Leaf explants, derived from plants regenerated from SR, were inoculated with Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pBI121, which contains the neomycin phosphotransferase II (NPTII) and beta-glucuronidase (GUS) genes as selectable and visual markers, respectively. After co-cultivation, the explants were selected on solidified MS medium with 0.5 mg/L benzylamino purine (BAP), 100 mg/L kanamycin and 250 mg/L cefotaxime. Kanamycin-resistant calli were transferred to liquid rooting medium. The newly regenerated, kanamycin-resistant roots were harvested and SR cultures re-established, which exhibited all the characteristics of the original SR. Furthermore, kanamycin-resistant roots cultured onto solidified MS medium supplemented with 0.5 mg/L BAP produced plants at the same rate as control SR. Six months after gene transfer, PCR analysis and histochemical locating indicated that the NPTII gene was integrated into the genome and that the GUS gene was regularly expressed in leaves, roots and nodules, respectively. The protocol makes it now possible to produce transformed SR and nodules as well as transgenic plants from transformed SR.

  15. Functional analysis of autophagy genes via Agrobacterium-mediated transformation in the vascular Wilt fungus Verticillium dahliae.

    PubMed

    Zhou, Lei; Zhao, Jun; Guo, Wangzhen; Zhang, Tianzhen

    2013-08-20

    Autophagy is a widely conserved intracellular process for degradation and recycling of proteins, organelles and cytoplasm in eukaryotic organisms and is now emerging as an important process in foliar infection by many plant pathogenic fungi. However, the role of autophagy in soil-borne fungal physiology and infection biology is poorly understood. Here, we report the establishment of an Agrobacterium tumefaciens-mediated transformation (ATMT) system and its application to investigate two autophagy genes, VdATG8 and VdATG12, by means of targeted gene replacement and complementation. Transformation of a cotton-infecting Verticillium dahliae strain Vd8 with a novel binary vector pCOM led to the production of 384 geneticin-resistant transformants per 1 × 10(6) conidia. V. dahliae mutants lacking either VdATG8 or VdATG12 exhibited reduced conidiation and impaired aerial hyphae production. Disease development on Arabidopsis plants was slightly delayed when inoculated with VdATG8 or VdATG12 gene deletion mutants, compared with the wild-type and gene complemented strains. Surprisingly, in vitro inoculation with unimpaired roots revealed that the abilities of root invasion were not affected in gene deletion mutants. These results indicate that autophagy is necessary for aerial hyphae development and plant colonization but not for root infection in V. dahliae.

  16. Increased Agrobacterium-mediated transformation and rooting efficiencies in canola (Brassica napus L.) from hypocotyl segment explants

    NASA Technical Reports Server (NTRS)

    Cardoza, V.; Stewart, C. N.

    2003-01-01

    An efficient protocol for the production of transgenic Brassica napus cv. Westar plants was developed by optimizing two important parameters: preconditioning time and co-cultivation time. Agrobacterium tumefaciens-mediated transformation was performed using hypocotyls as explant tissue. Two variants of a green fluorescent protein (GFP)-encoding gene--mGFP5-ER and eGFP--both under the constitutive expression of the cauliflower mosaic virus 35S promoter, were used for the experiments. Optimizing the preconditioning time to 72 h and co-cultivation time with Agrobacterium to 48 h provided the increase in the transformation efficiency from a baseline of 4% to 25%. With mGFP5-ER, the transformation rate was 17% and with eGFP it was 25%. Transgenic shoots were selected on 200 mg/l kanamycin. Rooting efficiency was 100% on half-strength Murashige and Skoog medium with 10 g/l sucrose and 0.5 mg/l indole butyric acid in the presence of kanamycin.

  17. Identifying a Carotenoid Cleavage Dioxygenase 4a Gene and Its Efficient Agrobacterium-Mediated Genetic Transformation in Bixa orellana L.

    PubMed

    Sankari, Mohan; Hemachandran, Hridya; Anantharaman, Amirtha; Babu, Subramanian; Madrid, Renata Rivera; C, George Priya Doss; Fulzele, Devanand P; Siva, Ramamoorthy

    2016-07-01

    Carotenoids are metabolized to apocarotenoids through the pathway catalysed by carotenoid cleavage oxygenases (CCOs). The apocarotenoids are economically important as it is known to have therapeutic as well as industrial applications. For instance, bixin from Bixa orellana and crocin from Crocus sativus are commercially used as a food colourant and cosmetics since prehistoric time. In our present study, CCD4a gene has been identified and isolated from leaves of B. orellana for the first time and named as BoCCD4a; phylogenetic analysis was carried out using CLUSTAL W. From sequence analysis, BoCCD4a contains two exons and one intron, which was compared with the selected AtCCD4, RdCCD4, GmCCD4 and CmCCD4a gene. Further, the BoCCD4a gene was cloned into pCAMBIA 1301, transformed into Agrobacterium tumefaciens EHA105 strain and subsequently transferred into hypocotyledons and callus of B. orellana by agro-infection. Selection of stable transformation was screened on the basis of PCR detection by using GUS and hptII specific primer, which was followed by histochemical characterization. The percent transient GUS expression in hypocotyledons and callus was 84.4 and 80 %, respectively. The expression of BoCCD4a gene in B. orellana was confirmed through RT-PCR analysis. From our results, the sequence analysis of BoCCD4a gene of B. orellana was closely related to the CsCCD4 gene of C. sativus, which suggests this gene may have a role in various processes such as fragrance, insect attractant and pollination.

  18. Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume blackgram (Vigna mungo L. Hepper).

    PubMed

    Sainger, Manish; Chaudhary, Darshna; Dahiya, Savita; Jaiwal, Ranjana; Jaiwal, Pawan K

    2015-10-01

    An efficient, rapid and direct multiple shoot regeneration system amenable to Agrobacterium-mediated transformation from primary leaf with intact petiole of blackgram (Vigna mungo) is established for the first time. The effect of the explant type and its age, type and concentration of cytokinin and auxin either alone or in combination and genotype on multiple shoot regeneration efficiency and frequency was optimized. The primary leaf explants with petiole excised from 4-day-old seedlings directly developed multiple shoots (an average of 10 shoots/ explant) from the cut ends of the petiole in 95 % of the cultures on MSB (MS salts and B5 vitamins) medium containing 1.0 μM 6-benzylaminopurine. Elongated (2-3 cm) shoots were rooted on MSB medium with 2.5 μM indole-butyric acid and resulted plantlets were hardened and established in soil, where they resumed growth and reached maturity with normal seed set. The regenerated plants were morphologically similar to seed-raised plants and required 8 weeks time from initiation of culture to establish them in soil. The regeneration competent cells present at the cut ends of petiole are fully exposed and are, thus, easily accessible to Agrobacterium, making this plant regeneration protocol amenable for the production of transgenic plants. The protocol was further successfully used to develop fertile transgenic plants of blackgram using Agrobacterium tumefaciens strain EHA 105 carrying a binary vector pCAMBIA2301 that contains a neomycin phosphotransferase gene (nptII) and a β-glucuronidase (GUS) gene (uidA) interrupted with an intron. The presence and integration of transgenes in putative T0 plants were confirmed by polymerase chain reaction (PCR) and Southern blot hybridization, respectively. The transgenes were inherited in Mendelian fashion in T1 progeny and a transformation frequency of 1.3 % was obtained. This protocol can be effectively used for transferring new traits in blackgram and other legumes for their

  19. Micro-shock Wave Assisted Plant Transformation

    NASA Astrophysics Data System (ADS)

    Gnanadhas, Divya Prakash; Datey, Akshay; Chakravortty, Dipshikha; Gopalan, Jagadeesh

    Genetically modified (GM) crops are developed by transforming the desired DNA to plant. There are various methods employed to achieve the required transformation in plants. Agrobacterium mediated transformation and Biolistics or particle bombardment method are the most commonly used methods.

  20. [Agrobacterium-mediated sunflower transformation (Helianthus annuus L.) in vitro and in Planta using strain of LBA4404 harboring binary vector pBi2E with dsRNA-suppressor proline dehydrogenase gene].

    PubMed

    Tishchenko, E N; Komisarenko, A G; Mikhal'skaia, S I; Sergeeva, L E; Adamenko, N I; Morgun, B V; Kochetov, A V

    2014-01-01

    To estimate the efficiency of proline dehydrogenase gene suppression towards increasing of sunflower (Helianthus annuus L.) tolerance level to water deficit and salinity, we employed strain LBA4404 harboring pBi2E with double-stranded RNA-suppressor, which were prepared on basis arabidopsis ProDH1 gene. The techniques of Agrobacterium-mediated transformation in vitro and in planta during fertilization sunflower have been proposed. There was shown the genotype-depended integration of T-DNA in sunflower genome. PCR-analysis showed that ProDH1 presents in genome of inbred lines transformed in planta, as well as in T1- and T2-generations. In trans-genic regenerants the essential accumulation of free L-proline during early stages of in vitro cultivation under normal conditions was shown. There was established the essential accumulation of free proline in transgenic regenerants during cultivation under lethal stress pressure (0.4 M mannitol and 2.0% sea water salts) and its decline upon the recovery period. These data are declared about effectiveness of suppression of sunflower ProDH and gene participation in processes connected with osmotolerance.

  1. A Dark Incubation Period Is Important for Agrobacterium-Mediated Transformation of Mature Internode Explants of Sweet Orange, Grapefruit, Citron, and a Citrange Rootstock

    PubMed Central

    Marutani-Hert, Mizuri; Bowman, Kim D.; McCollum, Greg T.; Mirkov, T. Erik; Evens, Terence J.; Niedz, Randall P.

    2012-01-01

    Background Citrus has an extended juvenile phase and trees can take 2–20 years to transition to the adult reproductive phase and produce fruit. For citrus variety development this substantially prolongs the time before adult traits, such as fruit yield and quality, can be evaluated. Methods to transform tissue from mature citrus trees would shorten the evaluation period via the direct production of adult phase transgenic citrus trees. Methodology/Principal Findings Factors important for promoting shoot regeneration from internode explants from adult phase citrus trees were identified and included a dark incubation period and the use of the cytokinin zeatin riboside. Transgenic trees were produced from four citrus types including sweet orange, citron, grapefruit, and a trifoliate hybrid using the identified factors and factor settings. Significance The critical importance of a dark incubation period for shoot regeneration was established. These results confirm previous reports on the feasibility of transforming mature tissue from sweet orange and are the first to document the transformation of mature tissue from grapefruit, citron, and a trifoliate hybrid. PMID:23082165

  2. Agrobacterium-mediated transformation of potato using PLRV-rep and PVY CP genes and assessment of replicase mediated resistance against natural infection of PLRV

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Replicase-and coat protein gene-mediated resistances against potato leafroll virus (PLRV) and potato virus Y (PVY), respectively, demonstrated to be an effective way of protecting potato against two major virus problems (PLRV & PVY) world-wide. Potato cultivar Desiree was transformed using Agrobacte...

  3. Agrobacterium-mediated transient MaFT expression in mulberry (Morus alba L.) leaves.

    PubMed

    Wu, Su-Li; Yang, Xiao-Bing; Liu, Li-Qun; Jiang, Tao; Wu, Hai; Su, Chao; Qian, Yong-Hua; Jiao, Feng

    2015-01-01

    To optimize Agrobacterium-mediated transient transformation assay in mulberry (Morus alba L.), various infiltration methods, Agrobacterium tumefaciens (A. tumefaciens) strains, and bacterial concentrations were tested in mulberry seedlings. Compared with LBA4404, GV3101 harboring pBE2133 plasmids presented stronger GUS signals at 3 days post infiltration using syringe. Recombinant plasmids pBE2133:GFP and pBE2133:GFP:MaFT were successfully constructed. Transient expression of MaFT:GFP protein was found in leaves, petiole (cross section), and shoot apical meristem (SAM) of mulberry according to the GFP signal. Moreover, MaFT:GFP mRNA was also detected in leaves and SAM via RT-PCR and qRT-PCR. An efficient transient transformation system could be achieved in mulberry seedlings by syringe using A. tumefaciens GV3101 at the OD600 of 0.5. The movement of MaFT expression from leaves to SAM might trigger the precocious flowering of mulberry.

  4. Design, Construction, and Validation of Artificial MicroRNA Vectors Using Agrobacterium-Mediated Transient Expression System.

    PubMed

    Bhagwat, Basdeo; Chi, Ming; Han, Dianwei; Tang, Haifeng; Tang, Guiliang; Xiang, Yu

    2016-01-01

    Artificial microRNA (amiRNA) technology utilizes microRNA (miRNA) biogenesis pathway to produce artificially selected small RNAs using miRNA gene backbone. It provides a feasible strategy for inducing loss of gene function, and has been applied in functional genomics study, improvement of crop quality and plant virus disease resistance. A big challenge in amiRNA applications is the unpredictability of silencing efficacy of the designed amiRNAs and not all constructed amiRNA candidates would be expressed effectively in plant cells. We and others found that high efficiency and specificity in RNA silencing can be achieved by designing amiRNAs with perfect or almost perfect sequence complementarity to their targets. In addition, we recently demonstrated that Agrobacterium-mediated transient expression system can be used to validate amiRNA constructs, which provides a simple, rapid and effective method to select highly expressible amiRNA candidates for stable genetic transformation. Here, we describe the methods for design of amiRNA candidates with perfect or almost perfect base-pairing to the target gene or gene groups, incorporation of amiRNA candidates in miR168a gene backbone by one step inverse PCR amplification, construction of plant amiRNA expression vectors, and assay of transient expression of amiRNAs in Nicotiana benthamiana through agro-infiltration, small RNA extraction, and amiRNA Northern blot.

  5. Selectable marker elimination in the T0 generation by Agrobacterium-mediated co-transformation involving Mungbean yellow mosaic virus TrAP as a non-conditional negative selectable marker and bar for transient positive selection.

    PubMed

    RamanaRao, Mangu Venkata; Veluthambi, Karuppannan

    2010-05-01

    Transient selection involving the bar gene and non-conditional negative selection against stable T-DNA integration through the use of the Mungbean yellow mosaic virus (MYMV) transcriptional activator protein gene (TrAP) were used in a novel co-transformation strategy to generate selectable marker gene (SMG)-eliminated transgenic tobacco plants in the T(0) generation itself. Two compatible binary plasmids, pCam-bar-TrAP-gus harbouring bar as an SMG and the MYMV TrAP gene as a non-conditional negative selectable marker, and pGA472 with the nptII gene as an unselected experimental gene of interest (GOI) were placed in the Agrobacterium tumefaciens strain EHA105 and used for co-transformation. Transient selection with 5 mg l(-1) phosphinothricin (PPT) for 2-4 weeks and subsequent establishment in a PPT-minus medium yielded 114 plants from 200 leaf discs. The unselected nptII gene was detected by Southern blot analysis in 13 plants, revealing a co-transformation efficiency of 11.5%. Five of these plants harboured only the nptII gene (GOI) and not the bar gene (SMG). Thus, SMG elimination was achieved in the T(0) generation itself in 4.4% (5/114) of plants, which were transiently selected for 2-4 weeks on PPT. MYMV TrAP, a non-conditional negative selectable marker, effectively reduced the recovery of plants with stable integration of the SMG (bar).

  6. Development of an efficient agrobacterium-mediated gene targeting system for rice and analysis of rice knockouts lacking granule-bound starch synthase (Waxy) and β1,2-xylosyltransferase.

    PubMed

    Ozawa, Kenjirou; Wakasa, Yuhya; Ogo, Yuko; Matsuo, Kouki; Kawahigashi, Hiroyuki; Takaiwa, Fumio

    2012-04-01

    We have developed a high-frequency method for Agrobacterium-mediated gene targeting by combining an efficient transformation system using rice suspension-cultured calli and a positive/negative selection system. Compared with the conventional transformation system using calli on solid medium, transformation using suspension-cultured calli resulted in a 5- to 10-fold increase in the number of resistant calli per weight of starting material after positive/negative selection. Homologous recombination occurred in about 1.5% of the positive/negative selected calli. To evaluate the efficacy of our method, we show in this report that knockout rice plants containing either a disrupted Waxy (granule-bound starch synthase) or a disrupted Xyl (β1,2-xylosyltransferase) gene can be easily obtained by homologous recombination. Study of gene function using homologous recombination in higher plants can now be considered routine work as a direct result of this technical advance.

  7. Transient plant transformation mediated by Agrobacterium tumefaciens: Principles, methods and applications.

    PubMed

    Krenek, Pavel; Samajova, Olga; Luptovciak, Ivan; Doskocilova, Anna; Komis, George; Samaj, Jozef

    2015-11-01

    Agrobacterium tumefaciens is widely used as a versatile tool for development of stably transformed model plants and crops. However, the development of Agrobacterium based transient plant transformation methods attracted substantial attention in recent years. Transient transformation methods offer several applications advancing stable transformations such as rapid and scalable recombinant protein production and in planta functional genomics studies. Herein, we highlight Agrobacterium and plant genetics factors affecting transfer of T-DNA from Agrobacterium into the plant cell nucleus and subsequent transient transgene expression. We also review recent methods concerning Agrobacterium mediated transient transformation of model plants and crops and outline key physical, physiological and genetic factors leading to their successful establishment. Of interest are especially Agrobacterium based reverse genetics studies in economically important crops relying on use of RNA interference (RNAi) or virus-induced gene silencing (VIGS) technology. The applications of Agrobacterium based transient plant transformation technology in biotech industry are presented in thorough detail. These involve production of recombinant proteins (plantibodies, vaccines and therapeutics) and effectoromics-assisted breeding of late blight resistance in potato. In addition, we also discuss biotechnological potential of recombinant GFP technology and present own examples of successful Agrobacterium mediated transient plant transformations.

  8. Use of Agrobacterium rhizogenes strain 18r12v and paromomycin selection for transformation of Brachypodium distachyon and Brachypodium sylvaticum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genetic transformation of monocot grasses is a resource intensive process, the quality and efficiency of which is dependent in part upon the method of DNA introduction, as well as the ability to effectively separate transformed from wildtype tissue. Agrobacterium-mediated transformation of Brac...

  9. In planta transformation method for T-DNA transfer in orchids

    NASA Astrophysics Data System (ADS)

    Semiarti, Endang; Purwantoro, Aziz; Mercuriani, Ixora S.; Anggriasari, Anida M.; Jang, Seonghoe; Suhandono, Sony; Machida, Yasunori; Machida, Chiyoko

    2014-03-01

    Transgenic plant technology is an efficient tool to study the function of gene(s) in plant. The most popular and widely used technique is Agrobacterium-mediated transformation in which cocultivation was done by immersing the plant tissues/organ in overnight bacterial cultured for about 30 minutes to one hour under in vitro condition. In this experiment, we developed more easier technique that omitted the in vitro step during cocultivation with Agrobacterium, namely in planta transformation method. Pollinaria (compact pollen mass of orchid) of Phalaenopsis amabilis and Spathoglottis plicata orchids were used as target explants that were immersed into bacterial culture for 30 minutes, then dried up the pollinaria, the transformed pollinaria was used to pollinate orchid flowers. The T-DNA used for this experiments were Ubipro∷PaFT/A. tumefaciens GV3101 for P. amabilis and MeEF1α2 pro∷GUS/ A. tumefaciens LBA 4404 for S.plicata. Seeds that were produced from pollinated flowers were grown onto 10 mg/l hygromicin containing NP (New Phalaenopsis) medium. The existance of transgene in putative transformant protocorm (developing orchid embryo) genome was confirmed using PCR with specific primers of either PaFT or GUS genes. Histochemical GUS assay was also performed to the putative transformants. The result showed that transformation frequencies were 2.1 % in P. amabilis, and 0,53% in S. plicata. These results indicates that in planta transformation method could be used for Agrobacterium-mediated genetic transformation, with advantage easier and more secure work from contaminants than that of the in vitro method.

  10. Exploration of new perspectives and limitations in Agrobacterium mediated gene transfer technology. Progress report, [June 1, 1992-- May 31, 1994

    SciTech Connect

    Marton, L.

    1994-12-31

    This report describes progress aimed at constructing gene-transfer technology for Nicotiana plumbaginifolia. Most actual effort as described herein has so far been directed at exploring new perspectives and limitations in Agrobacterium mediated gene transfer. Accomplishments are described using a core homologous gene targeting vector.

  11. Advances in Agrobacterium tumefaciens-mediated genetic transformation of graminaceous crops.

    PubMed

    Singh, Roshan Kumar; Prasad, Manoj

    2016-05-01

    Steady increase in global population poses several challenges to plant science research, including demand for increased crop productivity, grain yield, nutritional quality and improved tolerance to different environmental factors. Transgene-based approaches are promising to address these challenges by transferring potential candidate genes to host organisms through different strategies. Agrobacterium-mediated gene transfer is one such strategy which is well known for enabling efficient gene transfer in both monocot and dicots. Due to its versatility, this technique underwent several advancements including development of improved in vitro plant regeneration system, co-cultivation and selection methods, and use of hyper-virulent strains of Agrobacterium tumefaciens harbouring super-binary vectors. The efficiency of this method has also been enhanced by the use of acetosyringone to induce the activity of vir genes, silver nitrate to reduce the Agrobacterium-induced necrosis and cysteine to avoid callus browning during co-cultivation. In the last two decades, extensive efforts have been invested towards achieving efficient Agrobacterium-mediated transformation in cereals. Though high-efficiency transformation systems have been developed for rice and maize, comparatively lesser progress has been reported in other graminaceous crops. In this context, the present review discusses the progress made in Agrobacterium-mediated transformation system in rice, maize, wheat, barley, sorghum, sugarcane, Brachypodium, millets, bioenergy and forage and turf grasses. In addition, it also provides an overview of the genes that have been recently transferred to these graminaceous crops using Agrobacterium, bottlenecks in this technique and future possibilities for crop improvement.

  12. Establishment of an Agrobacterium-mediated Inoculation System for Cucumber Green Mottle Mosaic Virus

    PubMed Central

    Kang, Minji; Seo, Jang-Kyun; Song, Dami; Choi, Hong-Soo; Kim, Kook-Hyung

    2015-01-01

    The infectious full-length cDNA clones of Cucumber green mottle mosaic virus (CGMMV) isolates KW and KOM, which were isolated from watermelon and oriental melon, respectively, were constructed under the control of the cauliflower mosaic virus 35S promoter. We successfully inoculated Nicotiana benthamiana with the cloned CGMMV isolates KW and KOM by Agrobacterium-mediated infiltration. Virulence and symptomatic characteristics of the cloned CGMMV isolates KW and KOM were tested on several indicator plants. No obvious differences between two cloned isolates in disease development were observed on the tested indicator plants. We also determined full genome sequences of the cloned CGMMV isolates KW and KOM. Sequence comparison revealed that only four amino acids (at positions 228, 699, 1212, and 1238 of the replicase protein region) differ between the cloned isolates KW and KOM. A previous study reported that the isolate KOM could not infect Chenopodium amaranticolor, but the cloned KOM induced chlorotic spots on the inoculated leaves. When compared with the previously reported sequence of the original KOM isolate, the cloned KOM contained one amino acid mutation (Ala to Thr) at position 228 of the replicase protein, suggesting that this mutation might be responsible for induction of chlorotic spots on the inoculated leaves of C. amaranticolor. PMID:26674677

  13. Agrobacterium Mediated Transient Gene Silencing (AMTS) in Stevia rebaudiana: Insights into Steviol Glycoside Biosynthesis Pathway

    PubMed Central

    Guleria, Praveen; Yadav, Sudesh Kumar

    2013-01-01

    Background Steviol glycoside biosynthesis pathway has emerged as bifurcation from ent-kaurenoic acid, substrate of methyl erythritol phosphate pathway that also leads to gibberellin biosynthesis. However, the genetic regulation of steviol glycoside biosynthesis has not been studied. So, in present study RNA interference (RNAi) based Agrobacterium mediated transient gene silencing (AMTS) approach was followed. SrKA13H and three SrUGTs (SrUGT85C2, SrUGT74G1 and SrUGT76G1) genes encoding ent-kaurenoic acid-13 hydroxylase and three UDP glycosyltransferases of steviol glycoside biosynthesis pathway were silenced in Stevia rebaudiana to understand its molecular mechanism and association with gibberellins. Methodology/Principal Findings RNAi mediated AMTS of SrKA13H and three SrUGTs has significantly reduced the expression of targeted endogenous genes as well as total steviol glycoside accumulation. While gibberellins (GA3) content was significantly enhanced on AMTS of SrUGT85C2 and SrKA13H. Silencing of SrKA13H and SrUGT85C2 was found to block the metabolite flux of steviol glycoside pathway and shifted it towards GA3 biosynthesis. Further, molecular docking of three SrUGT proteins has documented highest affinity of SrUGT76G1 for the substrates of alternate pathways synthesizing steviol glycosides. This could be a plausible reason for maximum reduction in steviol glycoside content on silencing of SrUGT76G1 than other genes. Conclusions SrKA13H and SrUGT85C2 were identified as regulatory genes influencing carbon flux between steviol glycoside and gibberellin biosynthesis. This study has also documented the existence of alternate steviol glycoside biosynthesis route. PMID:24023961

  14. Cell wall biochemical alterations during Agrobacterium-mediated expression of haemagglutinin-based influenza virus-like vaccine particles in tobacco.

    PubMed

    Le Mauff, François; Loutelier-Bourhis, Corinne; Bardor, Muriel; Berard, Caroline; Doucet, Alain; D'Aoust, Marc-André; Vezina, Louis-Philippe; Driouich, Azeddine; Couture, Manon M-J; Lerouge, Patrice

    2017-03-01

    Influenza virus-like particles (VLPs) have been shown to induce a safe and potent immune response through both humoral and cellular responses. They represent promising novel influenza vaccines. Plant-based biotechnology allows for the large-scale production of VLPs of biopharmaceutical interest using different model organisms, including Nicotiana benthamiana plants. Through this platform, influenza VLPs bud from the plasma membrane and accumulate between the membrane and the plant cell wall. To design and optimize efficient production processes, a better understanding of the plant cell wall composition of infiltrated tobacco leaves is a major interest for the plant biotechnology industry. In this study, we have investigated the alteration of the biochemical composition of the cell walls of N. benthamiana leaves subjected to abiotic and biotic stresses induced by the Agrobacterium-mediated transient transformation and the resulting high expression levels of influenza VLPs. Results show that abiotic stress due to vacuum infiltration without Agrobacterium did not induce any detectable modification of the leaf cell wall when compared to non infiltrated leaves. In contrast, various chemical changes of the leaf cell wall were observed post-Agrobacterium infiltration. Indeed, Agrobacterium infection induced deposition of callose and lignin, modified the pectin methylesterification and increased both arabinosylation of RG-I side chains and the expression of arabinogalactan proteins. Moreover, these modifications were slightly greater in plants expressing haemagglutinin-based VLP than in plants infiltrated with the Agrobacterium strain containing only the p19 suppressor of silencing.

  15. Exploration of new perspectives and limitations in Agrobacterium-mediated gene transfer technology. Final report, June 1, 1992--May 31, 1995

    SciTech Connect

    Marton, L.

    1996-02-01

    Genetic manipulation of plants often involves the introduction of homologous or partly homologous genes. Ectropic introduction of homologous sequences into plant genomes may trigger epigenetic changes, making expression of the genes unpredictable. The main project objective was to examine the feasibility of using Agrobacterium-mediated gene transfer for homologous gene targeting in plants.

  16. Environment Control to Improve Recombinant Protein Yields in Plants Based on Agrobacterium-Mediated Transient Gene Expression.

    PubMed

    Fujiuchi, Naomichi; Matoba, Nobuyuki; Matsuda, Ryo

    2016-01-01

    Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: (1) recombinant protein content per unit biomass and (2) recombinant protein productivity per unit area-time at the end of the upstream production. Because environmental factors in the upstream production have impacts on these parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and postinoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Preinoculation environmental factors associated with planting density, light quality, and nutrient supply affect plant characteristics, such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, postinoculation environmental factors, such as temperature, light intensity, and humidity, have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the postinoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain high recombinant

  17. Environment Control to Improve Recombinant Protein Yields in Plants Based on Agrobacterium-Mediated Transient Gene Expression

    PubMed Central

    Fujiuchi, Naomichi; Matoba, Nobuyuki; Matsuda, Ryo

    2016-01-01

    Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: (1) recombinant protein content per unit biomass and (2) recombinant protein productivity per unit area–time at the end of the upstream production. Because environmental factors in the upstream production have impacts on these parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and postinoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Preinoculation environmental factors associated with planting density, light quality, and nutrient supply affect plant characteristics, such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, postinoculation environmental factors, such as temperature, light intensity, and humidity, have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the postinoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain high recombinant

  18. Transformation method and wave control

    NASA Astrophysics Data System (ADS)

    Chang, Zheng; Hu, Jin; Hu, Geng-Kai

    2010-12-01

    Transformation method provides an efficient way to control wave propagation by materials. The transformed relations for field and material during a transformation are essential to fulfill this method. We propose a systematic method to derive the transformed relations for a general physic process, the constraint conditions are obtained by considering geometrical and physical constraint during a mapping. The proposed method is applied to Navier's equation for elastodynamics, Helmholtz's equation for acoustic wave and Maxwell's equation for electromagnetic wave, the corresponding transformed relations are derived, which can be used in the framework of transformation method for wave control. We show that contrary to electromagnetic wave, the transformed relations are not uniquely determined for elastic wave and acoustic wave, so we have a freedom to choose them differently. Using the obtained transformed relations, we also provide some examples for device design, a concentrator for elastic wave, devices for illusion acoustic and illusion optics are conceived and validated by numerical simulations.

  19. Spike-dip transformation of Setaria viridis.

    PubMed

    Saha, Prasenjit; Blumwald, Eduardo

    2016-04-01

    Traditional method of Agrobacterium-mediated transformation through the generation of tissue culture had limited success for Setaria viridis, an emerging C4 monocot model. Here we present an efficient in planta method for Agrobacterium-mediated genetic transformation of S. viridis using spike dip. Pre-anthesis developing spikes were dipped into a solution of Agrobacterium tumefaciens strain AGL1 harboring the β-glucuronidase (GUS) reporter gene driven by the cauliflower mosaic virus 35S (CaMV35S) promoter to standardize and optimize conditions for transient as well as stable transformations. A transformation efficiency of 0.8 ± 0.1% was obtained after dipping of 5-day-old S3 spikes for 20 min in Agrobacterium cultures containing S. viridis spike-dip medium supplemented with 0.025% Silwet L-77 and 200 μm acetosyringone. Reproducibility of this method was demonstrated by generating stable transgenic lines expressing β-glucuronidase plus (GUSplus), green fluorescent protein (GFP) and Discosoma sp. red fluorescent protein (DsRed) reporter genes driven by either CaMV35S or intron-interrupted maize ubiquitin (Ubi) promoters from three S. viridis genotypes. Expression of these reporter genes in transient assays as well as in T1 stable transformed plants was monitored using histochemical, fluorometric GUS activity and fluorescence microscopy. Molecular analysis of transgenic lines revealed stable integration of transgenes into the genome, and inherited transgenes expressed in the subsequent generations. This approach provides opportunities for the high-throughput transformation and potentially facilitates translational research in a monocot model plant.

  20. Transformative Mixed Methods Research

    ERIC Educational Resources Information Center

    Mertens, Donna M.

    2010-01-01

    Paradigms serve as metaphysical frameworks that guide researchers in the identification and clarification of their beliefs with regard to ethics, reality, knowledge, and methodology. The transformative paradigm is explained and illustrated as a framework for researchers who place a priority on social justice and the furtherance of human rights.…

  1. Citrus transformation using juvenile tissue explants.

    PubMed

    Orbović, Vladimir; Grosser, Jude W

    2015-01-01

    The most frequently used method for production of citrus transgenic plants is via Agrobacterium-mediated transformation of tissues found on explants obtained from juvenile seedlings. Within the last decade and especially within the last 5-6 years, this robust method was employed to produce thousands of transgenic plants. With the newly applied screening methods that allow easier and faster detection of transgenic shoots, estimates of transformation rate for some cultivars have gone up making this approach even more attractive. Although adjustments have to be made regarding the (varietal) source of the starting material and Agrobacterium strain used in each experiment preformed, the major steps of this procedure have not changed significantly if at all. Transgenic citrus plants produced this way belong to cultivars of rootstocks, sweet oranges, grapefruits, mandarins, limes, and lemons.

  2. Transformation of carotenoid biosynthetic genes using a micro-cross section method in kiwifruit (Actinidia deliciosa cv. Hayward).

    PubMed

    Kim, Misun; Kim, Seong-Cheol; Song, Kwan Jeong; Kim, Ho Bang; Kim, In-Jung; Song, Eun-Young; Chun, Seung-Jong

    2010-12-01

    Genetic transformation using a micro-cross section (MCS) technique was conducted to improve the carotenoid content in kiwifruit (Actinidia deliciosa cv. Hayward). The introduced carotenoid biosynthetic genes include geranylgeranyl diphosphate synthase (GGPS), phytoene desaturase (PDS), ζ-carotene desaturase (ZDS), β-carotene hydroxylase (CHX), and phytoene synthase (PSY). The transformed explants were selected on half-strength MS medium containing 0.001 mg l(-1) of 2,4-D and 0.1 mg l(-1) of zeatin, either 5 mg l(-1) hygromycin or 25 mg l(-1) kanamycin, and 500 mg l(-1) cefotaxime. The genomic PCR, genomic Southern blot analysis, and RT-PCR were performed to confirm the integration and expression of the transgenes. The transformation efficiencies of either kanamycin- or hygromycin-resistant shoots ranged from 2.9 to 22.1% depending on the target genes, and from 2.9 to 24.2% depending on the reporter genes. The selection efficiencies ranged from 66.7 to 100% for the target genes and from 95.8 to 100% for the reporter genes. Changes of carotenoid content in the several PCR-positive plants were determined by UPLC analysis. As a result, transgenic plants expressing either GGPS or PSY increased about 1.2- to 1.3-fold in lutein or β-carotene content compared to non-transgenic plants. Our results suggest that the Agrobacterium-mediated transformation efficiency of kiwifruit can be greatly increased by this MCS method and that the carotenoid biosynthetic pathway can be modified in kiwifruit by genetic transformation. Our results further suggest that GGPS and PSY genes could be major target genes to increase carotenoid contents in kiwifruit.

  3. Obtaining transgenic plants using the bio-active beads method.

    PubMed

    Liu, Haibo; Kawabe, Akira; Matsunaga, Sachihiro; Murakawa, Tomoko; Mizukami, Atsushi; Yanagisawa, Masanobu; Nagamori, Eiji; Harashima, Satoshi; Kobayashi, Akio; Fukui, Kiichi

    2004-04-01

    Several methods of transformation are currently available for delivering exogenous DNA into animal and plant cells. In this study, a novel and efficient transformation system for DNA delivery/expression with a capacity to transport DNA of high molecular weight was developed. This system can overcome the shortcomings of traditional transformation methods such as Agrobacterium-mediated transformation, particle bombardment, and the electroporation method. The method developed in this study uses calcium alginate micro beads to immobilize DNA molecules in combination with polyethylene glycol treatment. In addition, it is simple and low-cost, and requires limited equipment. Using this method, we have successfully transformed tobacco plants, screening by kanamycin resistance. The transformed genes in the transformants were confirmed by PCR and Southern hybridization.

  4. Barley (Hordeum vulgare L.) transformation using embryogenic pollen cultures.

    PubMed

    Otto, Ingrid; Müller, Andrea; Kumlehn, Jochen

    2015-01-01

    The temperate cereal barley is grown as a source of food, feed, and malt. The development of a broad range of genetic resources and associated technologies in this species has helped to establish barley as the prime model for the other Triticeae cereals. The specific advantage of the transformation method presented here is that transgene homozygosity is attained in the same generation as the transgenic event occurred through the coupling of haploid technology with Agrobacterium-mediated transformation. Pollen is haploid and, following transformation, can be induced to regenerate into haploid plantlets, which can subsequently subjected to colchicine treatment to obtain diploid, genetically fixed plants. The routine application of the method based on the winter-type barley cultivar 'Igri' over a period of over 10 years has achieved an average yield of about two transgenic plants per donor spike. The whole procedure from pollen isolation to non-segregating transgenic, mature grain takes less than 12 months.

  5. Agrobacterium-mediated transformation of Mexican lime (Citrus aurantifolia Swingle) using optimized systems for epicotyls and cotelydons

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Epicotyl and internodal stem segments provide the predominantly used explants for regeneration of transgenic citrus plants following co-cultivation with Agrobacterium. Previous reports using epicotyls segments from Mexican lime have shown low affinity for Agrobacterium tumefaciens infection which re...

  6. Development of an efficient transformation method by Agrobacterium tumefaciens and high throughput spray assay to identify transgenic plants for woodland strawberry (Fragaria vesca) using NPTII selection.

    PubMed

    Pantazis, Christopher J; Fisk, Sarah; Mills, Kerri; Flinn, Barry S; Shulaev, Vladimir; Veilleux, Richard E; Dan, Yinghui

    2013-03-01

    KEY MESSAGE : We developed an efficient Agrobacterium -mediated transformation method using an Ac/Ds transposon tagging construct for F. vesca and high throughput paromomycin spray assay to identify its transformants for strawberry functional genomics. Genomic resources for Rosaceae species are now readily available, including the Fragaria vesca genome, EST sequences, markers, linkage maps, and physical maps. The Rosaceae Genomic Executive Committee has promoted strawberry as a translational genomics model due to its unique biological features and transformability for fruit trait improvement. Our overall research goal is to use functional genomic and metabolic approaches to pursue high throughput gene discovery in the diploid woodland strawberry. F. vesca offers several advantages of a fleshy fruit typical of most fruit crops, short life cycle (seed to seed in 12-16 weeks), small genome size (206 Mbb/C), small plant size, self-compatibility, and many seeds per plant. We have developed an efficient Agrobacterium tumefaciens-mediated strawberry transformation method using kanamycin selection, and high throughput paromomycin spray assay to efficiently identify transgenic strawberry plants. Using our kanamycin transformation method, we were able to produce up to 98 independent kanamycin resistant insertional mutant lines using a T-DNA construct carrying an Ac/Ds transposon Launchpad system from a single transformation experiment involving inoculation of 22 leaf explants of F. vesca accession 551572 within approx. 11 weeks (from inoculation to soil). Transgenic plants with 1-2 copies of a transgene were confirmed by Southern blot analysis. Using our paromomycin spray assay, transgenic F. vesca plants were rapidly identified within 10 days after spraying.

  7. Functional genomic analysis of cotton genes with agrobacterium-mediated virus-induced gene silencing.

    PubMed

    Gao, Xiquan; Shan, Libo

    2013-01-01

    Cotton (Gossypium spp.) is one of the most agronomically important crops worldwide for its unique textile fiber production and serving as food and feed stock. Molecular breeding and genetic engineering of useful genes into cotton have emerged as advanced approaches to improve cotton yield, fiber quality, and resistance to various stresses. However, the understanding of gene functions and regulations in cotton is largely hindered by the limited molecular and biochemical tools. Here, we describe the method of an Agrobacterium infiltration-based virus-induced gene silencing (VIGS) assay to transiently silence endogenous genes in cotton at 2-week-old seedling stage. The genes of interest could be readily silenced with a consistently high efficiency. To monitor gene silencing efficiency, we have cloned cotton GrCla1 from G. raimondii, a homolog gene of Arabidopsis Cloroplastos alterados 1 (AtCla1) involved in chloroplast development, and inserted into a tobacco rattle virus (TRV) binary vector pYL156. Silencing of GrCla1 results in albino phenotype on the newly emerging leaves, serving as a visual marker for silencing efficiency. To further explore the possibility of using VIGS assay to reveal the essential genes mediating disease resistance to Verticillium dahliae, a fungal pathogen causing severe Verticillium wilt in cotton, we developed a seedling infection assay to inoculate cotton seedlings when the genes of interest are silenced by VIGS. The method we describe here could be further explored for functional genomic analysis of cotton genes involved in development and various biotic and abiotic stresses.

  8. Efficient Agrobacterium tumefaciens-mediated transformation and regeneration of garlic (Allium sativum) immature leaf tissue.

    PubMed

    Kenel, Fernand; Eady, Colin; Brinch, Sheree

    2010-03-01

    Transgenic garlic (Allium sativum) plants have been recovered directly from immature leaf material by selective culture following Agrobacterium-mediated transformation. This method involved the use of a binary vector containing the mgfp-ER reporter gene and hpt selectable marker, and followed a similar protocol developed previously for the transformation of immature onion embryos. The choice of tissue and post-transformation selection procedure resulted in a large increase in recovery of transgenic plants compared with previously confirmed allium transformation protocols. The presence of transgenes in the genome of the plants was confirmed using Southern analysis. This improvement in frequency and the use of clonal commercial "Printanor" germplasm now makes possible the integration of useful agronomic and quality traits into this crop.

  9. Overexpression of several Arabidopsis histone genes increases Agrobacterium-medicated transformation and transgene expression in plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Arabidopsis histone H2A-1 is important for Agrobacterium-mediated plant transformation. Mutation of HTA1, the gene encoding histone H2A-1, in the rat5 mutant results in decreased T-(transferred) DNA integration into the plant genome, whereas over-expression of HTA1 increases transformation freq...

  10. Advances in Maize Transformation Technologies and Development of Transgenic Maize

    PubMed Central

    Yadava, Pranjal; Abhishek, Alok; Singh, Reeva; Singh, Ishwar; Kaul, Tanushri; Pattanayak, Arunava; Agrawal, Pawan K.

    2017-01-01

    Maize is the principal grain crop of the world. It is also the crop where genetic engineering has been employed to a great extent to improve its various traits. The ability to transform maize is a crucial step for application of gene technology in maize improvement. There have been constant improvements in the maize transformation technologies over past several years. The choice of genotype and the explant material to initiate transformation and the different types of media to be used in various stages of tissue culture can have significant impact on the outcomes of the transformation efforts. Various methods of gene transfer, like the particle bombardment, protoplast transformation, Agrobacterium-mediated, in planta transformation, etc., have been tried and improved over years. Similarly, various selection systems for retrieval of the transformants have been attempted. The commercial success of maize transformation and transgenic development is unmatched by any other crop so far. Maize transformation with newer gene editing technologies is opening up a fresh dimension in transformation protocols and work-flows. This review captures the various past and recent facets in improvement in maize transformation technologies and attempts to present a comprehensive updated picture of the current state of the art in this area. PMID:28111576

  11. Transformation of somatic embryos of Prunus incisa ‘February Pink’ with a visible reporter gene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An Agrobacterium-mediated transformation system was developed for the ornamental cherry species Prunus incisa. This system uses both an antibiotic resistance gene (NPTII) and a visible selectable marker, the green fluorescent protein (GFP), to select plants. Cells from leaf and root explants were tr...

  12. Genetic Transformation of Switchgrass

    NASA Astrophysics Data System (ADS)

    Xi, Yajun; Ge, Yaxin; Wang, Zeng-Yu

    Switchgrass (Panicum virgatum L.) is a highly productive warm-season C4 species that is being developed into a dedicated biofuel crop. This chapter describes a protocol that allows the generation of transgenic switchgrass plants by Agrobacterium tumefaciens-mediated transformation. Embryogenic calluses induced from caryopses or inflorescences were used as explants for inoculation with A. tumefaciens strain EHA105. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Calluses resistant to hygromycin were obtained after 5-6 weeks of selection. Soil-grown switchgrass plants were regenerated about 6 months after callus induction and Agrobacterium-mediated transformation.

  13. Isolation and functional characterization of the promoter of a DEAD-box helicase Psp68 using Agrobacterium-mediated transient assay.

    PubMed

    Banu, Sufara Akhter; Huda, Kazi Md Kamrul; Tuteja, Narendra

    2014-04-30

    Helicases are molecular motor proteins that perform a variety of cellular functions including transcription, translation, DNA replication and repair, RNA maturation, ribosome synthesis, nuclear export and splicing processes. The p68 is an evolutionarily conserved protein which plays pivotal roles in all aspect RNA metabolism processes. It is well established that helicases provides abiotic stress adaptation in plants but analysis of cis-regulatory elements present in the upstream regions is still infancy. Here we report isolation and functional characterization of the promoter of a DEAD-box helicase Psp68 in response to abiotic stress and hormonal regulation. The promoter of Psp68 was isolated by gene walking PCR from pea genomic DNA library constructed in BD genome walker kit. In silico analysis revealed that promoter of Psp68 contained a TATA, a CAAT motif and also harbors some important stress and hormone associated cis regulatory elements, including E-box, AGAAA, GATA-box, ACGT, GAAAA and GTCTC. Functional analyses were performed by Agrobacterium-mediated transient assay in tobacco leaves. Very high level of GUS activity was observed in agroinfiltrated tobacco leaves by the construct carrying the Psp68 promoter::GUS, subjected to abiotic stress and exogenous hormonal treatments. Stress-inducible nature of Psp68 promoter opens possibility for the study of the gene regulation under stress condition. Therefore, may be useful in the field of agriculture and biotechnology.

  14. Fourier Transform Methods. Chapter 4

    NASA Technical Reports Server (NTRS)

    Kaplan, Simon G.; Quijada, Manuel A.

    2015-01-01

    This chapter describes the use of Fourier transform spectrometers (FTS) for accurate spectrophotometry over a wide spectral range. After a brief exposition of the basic concepts of FTS operation, we discuss instrument designs and their advantages and disadvantages relative to dispersive spectrometers. We then examine how common sources of error in spectrophotometry manifest themselves when using an FTS and ways to reduce the magnitude of these errors. Examples are given of applications to both basic and derived spectrophotometric quantities. Finally, we give recommendations for choosing the right instrument for a specific application, and how to ensure the accuracy of the measurement results..

  15. Suppression of Ku70/80 or Lig4 leads to decreased stable transformation and enhanced homologous recombination in rice.

    PubMed

    Nishizawa-Yokoi, Ayako; Nonaka, Satoko; Saika, Hiroaki; Kwon, Yong-Ik; Osakabe, Keishi; Toki, Seiichi

    2012-12-01

    Evidence for the involvement of the nonhomologous end joining (NHEJ) pathway in Agrobacterium-mediated transferred DNA (T-DNA) integration into the genome of the model plant Arabidopsis remains inconclusive. Having established a rapid and highly efficient Agrobacterium-mediated transformation system in rice (Oryza sativa) using scutellum-derived calli, we examined here the involvement of the NHEJ pathway in Agrobacterium-mediated stable transformation in rice. Rice calli from OsKu70, OsKu80 and OsLig4 knockdown (KD) plants were infected with Agrobacterium harboring a sensitive emerald luciferase (LUC) reporter construct to evaluate stable expression and a green fluorescent protein (GFP) construct to monitor transient expression of T-DNA. Transient expression was not suppressed, but stable expression was reduced significantly, in KD plants. Furthermore, KD-Ku70 and KD-Lig4 calli exhibited an increase in the frequency of homologous recombination (HR) compared with control calli. In addition, suppression of OsKu70, OsKu80 and OsLig4 induced the expression of HR-related genes on treatment with DNA-damaging agents. Our findings suggest strongly that NHEJ is involved in Agrobacterium-mediated stable transformation in rice, and that there is a competitive and complementary relationship between the NHEJ and HR pathways for DNA double-strand break repair in rice.

  16. Floral-Dip Transformation of Flax (Linum usitatissimum) to Generate Transgenic Progenies with a High Transformation Rate

    PubMed Central

    Bastaki, Nasmah K.; Cullis, Christopher A.

    2014-01-01

    Agrobacterium-mediated plant transformation via floral-dip is a widely used technique in the field of plant transformation and has been reported to be successful for many plant species. However, flax (Linum usitatissimum) transformation by floral-dip has not been reported. The goal of this protocol is to establish that Agrobacterium and the floral-dip method can be used to generate transgenic flax. We show that this technique is simple, inexpensive, efficient, and more importantly, gives a higher transformation rate than the current available methods of flax transformation. In summary, inflorescences of flax were dipped in a solution of Agrobacterium carrying a binary vector plasmid (T-DNA fragment plus the Linum Insertion Sequence, LIS-1) for 1 - 2 min. The plants were laid flat on their side for 24 hr. Then, plants were maintained under normal growth conditions until the next treatment. The process of dipping was repeated 2 - 3 times, with approximately 10 - 14 day intervals between dipping. The T1 seeds were collected and germinated on soil. After approximately two weeks, treated progenies were tested by direct PCR; 2 - 3 leaves were used per plant plus the appropriate T-DNA primers. Positive transformants were selected and grown to maturity. The transformation rate was unexpectedly high, with 50 - 60% of the seeds from treated plants being positive transformants. This is a higher transformation rate than those reported for Arabidopsis thaliana and other plant species, using floral-dip transformation. It is also the highest, which has been reported so far, for flax transformation using other methods for transformation. PMID:25549243

  17. Physical methods for genetic plant transformation

    NASA Astrophysics Data System (ADS)

    Rivera, Ana Leonor; Gómez-Lim, Miguel; Fernández, Francisco; Loske, Achim M.

    2012-09-01

    Production of transgenic plants is a routine process for many crop species. Transgenes are introduced into plants to confer novel traits such as improved nutritional qualities, tolerance to pollutants, resistance to pathogens and for studies of plant metabolism. Nowadays, it is possible to insert genes from plants evolutionary distant from the host plant, as well as from fungi, viruses, bacteria and even animals. Genetic transformation requires penetration of the transgene through the plant cell wall, facilitated by biological or physical methods. The objective of this article is to review the state of the art of the physical methods used for genetic plant transformation and to describe the basic physics behind them.

  18. Parallel algorithms for the spectral transform method

    SciTech Connect

    Foster, I.T.; Worley, P.H.

    1997-05-01

    The spectral transform method is a standard numerical technique for solving partial differential equations on a sphere and is widely used in atmospheric circulation models. Recent research has identified several promising algorithms for implementing this method on massively parallel computers; however, no detailed comparison of the different algorithms has previously been attempted. In this paper, the authors describe these different parallel algorithms and report on computational experiments that they have conducted to evaluate their efficiency on parallel computers. The experiments used a testbed code that solves the nonlinear shallow water equations on a sphere; considerable care was taken to ensure that the experiments provide a fair comparison of the different algorithms and that the results are relevant to global models. The authors focus on hypercube- and mesh-connected multicomputers with cut-through routing, such as the Intel iPSC/860, DELTA, and Paragon, and the nCUBE/2, but they also indicate how the results extend to other parallel computer architectures. The results of this study are relevant not only to the spectral transform method but also to multidimensional fast Fourier transforms (FFTs) and other parallel transforms.

  19. Discrete fourier transform (DFT) analysis for applications using iterative transform methods

    NASA Technical Reports Server (NTRS)

    Dean, Bruce H. (Inventor)

    2012-01-01

    According to various embodiments, a method is provided for determining aberration data for an optical system. The method comprises collecting a data signal, and generating a pre-transformation algorithm. The data is pre-transformed by multiplying the data with the pre-transformation algorithm. A discrete Fourier transform of the pre-transformed data is performed in an iterative loop. The method further comprises back-transforming the data to generate aberration data.

  20. Fourier transform methods in local gravity modeling

    NASA Technical Reports Server (NTRS)

    Harrison, J. C.; Dickinson, M.

    1989-01-01

    New algorithms were derived for computing terrain corrections, all components of the attraction of the topography at the topographic surface and the gradients of these attractions. These algoriithms utilize fast Fourier transforms, but, in contrast to methods currently in use, all divergences of the integrals are removed during the analysis. Sequential methods employing a smooth intermediate reference surface were developed to avoid the very large transforms necessary when making computations at high resolution over a wide area. A new method for the numerical solution of Molodensky's problem was developed to mitigate the convergence difficulties that occur at short wavelengths with methods based on a Taylor series expansion. A trial field on a level surface is continued analytically to the topographic surface, and compared with that predicted from gravity observations. The difference is used to compute a correction to the trial field and the process iterated. Special techniques are employed to speed convergence and prevent oscillations. Three different spectral methods for fitting a point-mass set to a gravity field given on a regular grid at constant elevation are described. Two of the methods differ in the way that the spectrum of the point-mass set, which extends to infinite wave number, is matched to that of the gravity field which is band-limited. The third method is essentially a space-domain technique in which Fourier methods are used to solve a set of simultaneous equations.

  1. Ensemble transform sensitivity method for adaptive observations

    NASA Astrophysics Data System (ADS)

    Zhang, Yu; Xie, Yuanfu; Wang, Hongli; Chen, Dehui; Toth, Zoltan

    2016-01-01

    The Ensemble Transform (ET) method has been shown to be useful in providing guidance for adaptive observation deployment. It predicts forecast error variance reduction for each possible deployment using its corresponding transformation matrix in an ensemble subspace. In this paper, a new ET-based sensitivity (ETS) method, which calculates the gradient of forecast error variance reduction in terms of analysis error variance reduction, is proposed to specify regions for possible adaptive observations. ETS is a first order approximation of the ET; it requires just one calculation of a transformation matrix, increasing computational efficiency (60%-80% reduction in computational cost). An explicit mathematical formulation of the ETS gradient is derived and described. Both the ET and ETS methods are applied to the Hurricane Irene (2011) case and a heavy rainfall case for comparison. The numerical results imply that the sensitive areas estimated by the ETS and ET are similar. However, ETS is much more efficient, particularly when the resolution is higher and the number of ensemble members is larger.

  2. Parallel algorithms for the spectral transform method

    SciTech Connect

    Foster, I.T.; Worley, P.H.

    1994-04-01

    The spectral transform method is a standard numerical technique for solving partial differential equations on a sphere and is widely used in atmospheric circulation models. Recent research has identified several promising algorithms for implementing this method on massively parallel computers; however, no detailed comparison of the different algorithms has previously been attempted. In this paper, we describe these different parallel algorithms and report on computational experiments that we have conducted to evaluate their efficiency on parallel computers. The experiments used a testbed code that solves the nonlinear shallow water equations or a sphere; considerable care was taken to ensure that the experiments provide a fair comparison of the different algorithms and that the results are relevant to global models. We focus on hypercube- and mesh-connected multicomputers with cut-through routing, such as the Intel iPSC/860, DELTA, and Paragon, and the nCUBE/2, but also indicate how the results extend to other parallel computer architectures. The results of this study are relevant not only to the spectral transform method but also to multidimensional FFTs and other parallel transforms.

  3. Improved genetic transformation of cork oak (Quercus suber L.).

    PubMed

    Alvarez-Fernández, Rubén; Ordás, Ricardo-Javier

    2012-01-01

    An Agrobacterium-mediated transformation system for selected mature Quercus suber L. trees has been established. Leaf-derived somatic embryos in an early stage of development are inoculated with an AGL1 strain harboring a kanamycin-selectable plasmid carrying the gene of interest. The transformed embryos are induced to germinate and the plantlets transferred to soil. This protocol, from adult cork oak to transformed plantlet, can be completed in about one and a half years. Transformation efficiencies (i.e., percentage of inoculated explants that yield independent transgenic embryogenic lines) vary depending on the cork oak genotype, reaching up to 43%.

  4. Drought-tolerant rice germplasm developed from an Oryza officinalis transformation-competent artificial chromosome clone.

    PubMed

    Liu, R; Zhang, H H; Chen, Z X; Shahid, M Q; Fu, X L; Liu, X D

    2015-10-29

    Oryza officinalis has proven to be a natural gene reservoir for the improvement of domesticated rice as it carries many desirable traits; however, the transfer of elite genes to cultivated rice by conventional hybridization has been a challenge for rice breeders. In this study, the conserved sequence of plant stress-related NAC transcription factors was selected as a probe to screen the O. officinalis genomic transformation-competent artificial chromosome library by Southern blot; 11 positive transformation-competent artificial chromosome clones were subsequently detected. By Agrobacterium-mediated transformation, an indica rice variety, Huajingxian 74 (HJX74), was transformed with a TAC clone harboring a NAC gene-positive genomic fragment from O. officinalis. Molecular analysis revealed that the O. officinalis genomic fragment was integrated into the genome of HJX74. The transgenic lines exhibited high tolerance to drought stress. Our results demonstrate that the introduction of stress-related transformation-competent artificial chromosome clones, coupled with a transgenic validation approach, is an effective method of transferring agronomically important genes from O. officinalis to cultivated rice.

  5. Analysis method for Fourier transform spectroscopy

    NASA Technical Reports Server (NTRS)

    Park, J. H.

    1983-01-01

    A fast Fourier transform technique is given for the simulation of those distortion effects in the instrument line shape of the interferometric spectrum that are due to errors in the measured interferogram. The technique is applied to analyses of atmospheric absorption spectra and laboratory spectra. It is shown that the nonlinear least squares method can retrieve the correct information from the distorted spectrum. Analyses of HF absorption spectra obtained in a laboratory and solar CO absorption spectra gathered by a balloon-borne interferometer indicate that the retrieved amount of absorbing gas is less than the correct value in most cases, if the interferogram distortion effects are not included in the analysis.

  6. Transformation of radish (Raphanus sativus L.) via sonication and vacuum infiltration of germinated seeds with Agrobacterium harboring a group 3 LEA gene from B. napus.

    PubMed

    Park, Byong-Jin; Liu, Zaochang; Kanno, Akira; Kameya, Toshiaki

    2005-10-01

    A protocol for producing transgenic radish (Raphanus sativus) was obtained by using both ultrasonic and vacuum infiltration assisted, Agrobacterium-mediated transformation. The Agrobacterium strain LBA4404 contained the binary vector pBI121-LEA (late embyogenesis abundant), which carried a Group 3 LEA gene, from Brassica napus. Among six combinations, Agrobacterium-mediated transformation assisted by a combination of 5-min sonication with 5-min vacuum infiltration resulted in the highest transformation frequency. The existence, integration and expression of transferred LEA gene in transgenic T(1) plants were confirmed by PCR, genomic Southern and Western blot analysis. Transgenic radish demonstrated better growth performance than non-transformed control plants under osmotic and salt stress conditions. Accumulation of Group 3 LEA protein in the vegetative tissue of transgenic radish conferred increased tolerance to water deficit and salt stress.

  7. Apparatus and method for transforming living cells

    DOEpatents

    Okandan, Murat; Galambos, Paul C.

    2003-11-11

    An apparatus and method are disclosed for in vitro transformation of living cells. The apparatus, which is formed as a microelectromechanical device by surface micromachining, can be used to temporarily disrupt the cell walls or membrane of host cells one at a time so that a particular substance (e.g. a molecular tag, nucleic acid, bacteria, virus etc.) can be introduced into the cell. Disruption of the integrity of the host cells (i.e. poration) can be performed mechanically or electrically, or by both while the host cells are contained within a flow channel. Mechanical poration is possible using a moveable member which has a pointed or serrated edge and which is driven by an electrostatic actuator to abrade, impact or penetrate the host cell. Electroporation is produced by generating a relatively high electric field across the host cell when the host cell is located in the flow channel between a pair of electrodes having a voltage applied therebetween.

  8. Use of Agrobacterium rhizogenes Strain 18r12v and Paromomycin Selection for Transformation of Brachypodium distachyon and Brachypodium sylvaticum

    PubMed Central

    Collier, Ray; Bragg, Jennifer; Hernandez, Bryan T.; Vogel, John P.; Thilmony, Roger

    2016-01-01

    The genetic transformation of monocot grasses is a resource intensive process, the quality and efficiency of which is dependent in part upon the method of DNA introduction, as well as the ability to effectively separate transformed from wildtype tissue. Agrobacterium-mediated transformation of Brachypodium has relied mainly on Agrobacterium tumefaciens strain AGL1. Currently the antibiotic hygromycin B has been the selective agent of choice for robust identification of transgenic calli in Brachypodium distachyon and Brachypodium sylvaticum but few other chemicals have been shown to work as well for selection of transgenic Brachypodium cells in tissue culture. This study demonstrates that Agrobacterium rhizogenes strain 18r12v and paromomycin selection can be successfully used for the efficient generation of transgenic B. distachyon and B. sylvaticum. Additionally we observed that the transformation rates were similar to or higher than those obtained with A. tumefaciens strain AGL1 and hygromycin selection. The A. rhizogenes strain 18r12v harboring the pARS1 binary vector and paromomycin selection is an effective means of generating transgenic Brachypodium plants. This novel approach will facilitate the transgenic complementation of T-DNA knockout mutants of B. distachyon which were created using hygromycin selection, as well as aid the implementation of more complex genome manipulation strategies which require multiple rounds of transformation. PMID:27252729

  9. Use of Agrobacterium rhizogenes strain 18r12v and paromomycin selection for transformation of Brachypodium distachyon and Brachypodium sylvaticum

    DOE PAGES

    Collier, Ray; Bragg, Jennifer; Hernandez, Bryan T.; ...

    2016-05-24

    In this study, the genetic transformation of monocot grasses is a resource intensive process, the quality and efficiency of which is dependent in part upon the method of DNA introduction, as well as the ability to effectively separate transformed from wildtype tissue. Agrobacterium-mediated transformation of Brachypodium has relied mainly on Agrobacterium tumefaciens strain AGL1. Currently the antibiotic hygromycin B has been the selective agent of choice for robust identification of transgenic calli in Brachypodium distachyon and Brachypodium sylvaticum but few other chemicals have been shown to work as well for selection of transgenic Brachypodium cells in tissue culture. This studymore » demonstrates that Agrobacterium rhizogenes strain 18r12v and paromomycin selection can be successfully used for the efficient generation of transgenic B. distachyon and B. sylvaticurn. Additionally we observed that the transformation rates were similar to or higher than those obtained with A. turnefaciens strain AGL1 and hygromycin selection. The A. rhizogenes strain 18r12v harboring the pARS1 binary vector and paromomycin selection is an effective means of generating transgenic Brachypodium plants. This novel approach will facilitate the transgenic complementation of T-DNA knockout mutants of B. distachyon which were created using hygromycin selection, as well as aid the implementation of more complex genome manipulation strategies which require multiple rounds of transformation.« less

  10. Agrobacterium tumefaciens-mediated transformation of taro (Colocasia esculenta (L.) Schott) with a rice chitinase gene for improved tolerance to a fungal pathogen Sclerotium rolfsii.

    PubMed

    He, Xiaoling; Miyasaka, Susan C; Fitch, Maureen M M; Moore, Paul H; Zhu, Yun J

    2008-05-01

    Taro (Colocasia esculenta) is one of the most important crops in the Pacific Islands, however, taro yields have been declining in Hawaii over the past 30 years partly due to diseases caused by oomycete and fungal pathogens. In this study, an efficient Agrobacterium tumefaciens-mediated transformation method for taro is first reported. In total, approximately 200 pieces (8 g) of embryogenic calluses were infected with the super-virulent A. tumefaciens strain EHA105 harboring the plant transformation plasmid pBI121/ricchi11 that contains the rice chitinase gene ricchi11. The presence and expression of the transgene ricchi11 in six independent transgenic lines was confirmed using polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR). Southern blot analysis of the six independent lines indicated that three out of six (50%) had integrated a single copy of the transgene, and the other three lines had two or three copies of the transgene. Compared to the particle bombardment transformation of taro method, which was used in the previous studies, the Agrobacterium-mediated transformation method obtained 43-fold higher transformation efficiency. In addition, these six transgenic lines via Agrobacterium may be more effective for transgene expression as a result of single-copy or low-copy insertion of the transgene than the single line with multiple copies of the transgene via particle bombardment. In a laboratory bioassay, all six transgenic lines exhibited increased tolerance to the fungal pathogen Sclerotium rolfsii, ranging from 42 to 63% reduction in lesion expansion.

  11. Cloning of TPS gene from eelgrass species Zostera marina and its functional identification by genetic transformation in rice.

    PubMed

    Zhao, Feng; Li, Qiuying; Weng, Manli; Wang, Xiuliang; Guo, Baotai; Wang, Li; Wang, Wei; Duan, Delin; Wang, Bin

    2013-12-01

    The full-length cDNA sequence (2613 bp) of the trehalose-6-phosphate synthase (TPS) gene of eelgrass Zostera marina (ZmTPS) was identified and cloned. Z. marina is a kind of seed-plant growing in sea water during its whole life history. The open reading frame (ORF) region of ZmTPS gene encodes a protein of 870 amino acid residues and a stop codon. The corresponding genomic DNA sequence is 3770 bp in length, which contains 3 exons and 2 introns. The ZmTPS gene was transformed into rice variety ZH11 via Agrobacterium-mediated transformation method. After antibiotic screening, molecular characterization, salt-tolerance and trehalose content determinations, two transgenic lines resistant to 150 mM NaCL solutions were screened. Our study results indicated that the ZmTPS gene was integrated into the genomic DNA of the two transgenic rice lines and could be expressed well. Moreover, the detection of the transformed ZmTPS gene in the progenies of the two transgenic lines was performed from T1 to T4 generations; and results suggested that the transformed ZmTPS gene can be transmitted from parent to the progeny in transgenic rice.

  12. Transformative, Mixed Methods Checklist for Psychological Research with Mexican Americans

    ERIC Educational Resources Information Center

    Canales, Genevieve

    2013-01-01

    This is a description of the creation of a research methods tool, the "Transformative, Mixed Methods Checklist for Psychological Research With Mexican Americans." For conducting literature reviews of and planning mixed methods studies with Mexican Americans, it contains evaluative criteria calling for transformative mixed methods, perspectives…

  13. Agrobacterium-mediated disruption of a nonribosomal peptide synthetase gene in the invertebrate pathogen Metarhizium anisopliae reveals a peptide spore factor

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Numerous secondary metabolites have been isolated from the insect pathogenic fungus Metarhizium anisopliae, but the roles of these compounds as virulence factors in disease development are poorly understood. We targeted for disruption by Agrobacterium tumefaciens-mediated transformation a putative n...

  14. Hough transform method for track finding in center drift chamber

    NASA Astrophysics Data System (ADS)

    Azmi, K. A. Mohammad Kamal; Wan Abdullah, W. A. T.; Ibrahim, Zainol Abidin

    2016-01-01

    Hough transform is a global tracking method used which had been expected to be faster approach for tracking the circular pattern of electron moving in Center Drift Chamber (CDC), by transforming the point of hit into a circular curve. This paper present the implementation of hough transform method for the reconstruction of tracks in Center Drift Chamber (CDC) which have been generated by random number in C language programming. Result from implementation of this method shows higher peak of circle parameter value (xc,yc,rc) that indicate the similarity value of the parameter needed for circular track in CDC for charged particles in the region of CDC.

  15. Hough transform method for track finding in center drift chamber

    SciTech Connect

    Azmi, K. A. Mohammad Kamal Wan Abdullah, W. A. T. Ibrahim, Zainol Abidin

    2016-01-22

    Hough transform is a global tracking method used which had been expected to be faster approach for tracking the circular pattern of electron moving in Center Drift Chamber (CDC), by transforming the point of hit into a circular curve. This paper present the implementation of hough transform method for the reconstruction of tracks in Center Drift Chamber (CDC) which have been generated by random number in C language programming. Result from implementation of this method shows higher peak of circle parameter value (xc,yc,rc) that indicate the similarity value of the parameter needed for circular track in CDC for charged particles in the region of CDC.

  16. Anisotropy minimization via least squares method for transformation optics.

    PubMed

    Junqueira, Mateus A F C; Gabrielli, Lucas H; Spadoti, Danilo H

    2014-07-28

    In this work the least squares method is used to reduce anisotropy in transformation optics technique. To apply the least squares method a power series is added on the coordinate transformation functions. The series coefficients were calculated to reduce the deviations in Cauchy-Riemann equations, which, when satisfied, result in both conformal transformations and isotropic media. We also present a mathematical treatment for the special case of transformation optics to design waveguides. To demonstrate the proposed technique a waveguide with a 30° of bend and with a 50% of increase in its output width was designed. The results show that our technique is simultaneously straightforward to be implement and effective in reducing the anisotropy of the transformation for an extremely low value close to zero.

  17. Direct Linear Transformation Method for Three-Dimensional Cinematography

    ERIC Educational Resources Information Center

    Shapiro, Robert

    1978-01-01

    The ability of Direct Linear Transformation Method for three-dimensional cinematography to locate points in space was shown to meet the accuracy requirements associated with research on human movement. (JD)

  18. Construction and characterization of a plant transformation-competent BIBAC library of the black Sigatoka-resistant banana Musa acuminata cv. Tuu Gia (AA).

    PubMed

    Ortiz-Vázquez, E; Kaemmer, D; Zhang, H-B; Muth, J; Rodríguez-Mendiola, M; Arias-Castro, C; James, Andrew

    2005-02-01

    A plant transformation-competent binary bacterial artificial chromosome (BIBAC) library was constructed from Musa acuminata cv. Tuu Gia (AA), a black Sigatoka-resistant diploid banana. After digestion of high-molecular-weight banana DNA by HindIII, several methods of DNA size selection were tested, followed by ligation, using a vector/insert molar ratio of 4:1. The library consists of 30,700 clones stored in 80 384-well microtiter plates. The mean insert size was estimated to be 100 kb, and the frequency of inserts with internal NotI sites was 61%. The majority of insert sizes fell into the range of 100+/-20 kb, making them suitable for Agrobacterium-mediated transformation. Only 1% and 0.9% of the clones contain chloroplast and mitochondrial DNA, respectively. This is the first BIBAC library for banana, estimated to represent five times its haploid genome (600 Mbp). It was demonstrated by hybridization that the library contains typical members of resistance gene and defense gene families that can be used for transformation of disease susceptible banana cultivars for banana genetic improvement.

  19. Cell Phones Transform a Science Methods Course

    ERIC Educational Resources Information Center

    Madden, Lauren

    2012-01-01

    A science methods instructor intentionally encouraged cell phone use for class work to discover how cell phones can be used as research tools to enhance the content and engage the students. The anecdotal evidence suggested that students who used their smartphones as research tools experienced the science content and pedagogical information…

  20. Data transformation methods for multiplexed assays

    DOEpatents

    Tammero, Lance F. Bentley; Dzenitis, John M; Hindson, Benjamin J

    2013-07-23

    Methods to improve the performance of an array assay are described. A correlation between fluorescence intensity-related parameters and negative control values of the assay is determined. The parameters are then adjusted as a function of the correlation. As a result, sensitivity of the assay is improved without changes in its specificity.

  1. Analytic solution for Telegraph equation by differential transform method

    NASA Astrophysics Data System (ADS)

    Biazar, J.; Eslami, M.

    2010-06-01

    In this article differential transform method (DTM) is considered to solve Telegraph equation. This method is a powerful tool for solving large amount of problems (Zhou (1986) [1], Chen and Ho (1999) [2], Jang et al. (2001) [3], Kangalgil and Ayaz (2009) [4], Ravi Kanth and Aruna (2009) [5], Arikoglu and Ozkol (2007) [6]). Using differential transform method, it is possible to find the exact solution or a closed approximate solution of an equation. To illustrate the ability and reliability of the method some examples are provided. The results reveal that the method is very effective and simple.

  2. Non-uniform beam vibration using Differential Transform Method

    NASA Astrophysics Data System (ADS)

    Shali, S.; Nagaraja, S. R.; Jafarali, P.

    2016-09-01

    The paper focuses on the vibration characteristics of non-uniform Euler- Bernoulli beam using Differential Transform Method (DTM). DTM is a numerical method to solve differential equations where the governing equations are reduced into a set of polynomials. Non-uniformity is considered corresponding to linear variation in breadth and height of the beam. The effect of taper ratio on the fundamental frequency of tapered beams is also analysed. The method has proved to be accurate, simple and effective for eigenvalue analysis. For the two cases of non-uniform beam analysed, the frequency computed by the method of differential transform is found to be comparable with the previously available results.

  3. SFCVQ and EZW coding method based on Karhunen-Loeve transformation and integer wavelet transformation

    NASA Astrophysics Data System (ADS)

    Yan, Jingwen; Chen, Jiazhen

    2007-03-01

    A new hyperspectral image compression method of spectral feature classification vector quantization (SFCVQ) and embedded zero-tree of wavelet (EZW) based on Karhunen-Loeve transformation (KLT) and integer wavelet transformation is represented. In comparison with the other methods, this method not only keeps the characteristics of high compression ratio and easy real-time transmission, but also has the advantage of high computation speed. After lifting based integer wavelet and SFCVQ coding are introduced, a system of nearly lossless compression of hyperspectral images is designed. KLT is used to remove the correlation of spectral redundancy as one-dimensional (1D) linear transform, and SFCVQ coding is applied to enhance compression ratio. The two-dimensional (2D) integer wavelet transformation is adopted for the decorrelation of 2D spatial redundancy. EZW coding method is applied to compress data in wavelet domain. Experimental results show that in comparison with the method of wavelet SFCVQ (WSFCVQ), the method of improved BiBlock zero tree coding (IBBZTC) and the method of feature spectral vector quantization (FSVQ), the peak signal-to-noise ratio (PSNR) of this method can enhance over 9 dB, and the total compression performance is improved greatly.

  4. A novel method for heterocyclic amide-thioamide transformations.

    PubMed

    Fathalla, Walid; Ali, Ibrahim A I; Pazdera, Pavel

    2017-01-01

    In this paper, we introduce a novel and convenient method for the transformation of heterocyclic amides into heteocyclic thioamides. A two-step approach was applied for this transformation: Firstly, we applied a chlorination of the heterocyclic amides to afford the corresponding chloroheterocycles. Secondly, the chloroherocycles and N-cyclohexyl dithiocarbamate cyclohexylammonium salt were heated in chloroform for 12 h at 61 °C to afford heteocyclic thioamides in excellent yields.

  5. Transformation-optics simulation method for stimulated Brillouin scattering

    NASA Astrophysics Data System (ADS)

    Zecca, Roberto; Bowen, Patrick T.; Smith, David R.; Larouche, Stéphane

    2016-12-01

    We develop an approach to enable the full-wave simulation of stimulated Brillouin scattering and related phenomena in a frequency-domain, finite-element environment. The method uses transformation-optics techniques to implement a time-harmonic coordinate transform that reconciles the different frames of reference used by electromagnetic and mechanical finite-element solvers. We show how this strategy can be successfully applied to bulk and guided systems, comparing the results with the predictions of established theory.

  6. A novel method for heterocyclic amide–thioamide transformations

    PubMed Central

    Ali, Ibrahim A I; Pazdera, Pavel

    2017-01-01

    In this paper, we introduce a novel and convenient method for the transformation of heterocyclic amides into heteocyclic thioamides. A two-step approach was applied for this transformation: Firstly, we applied a chlorination of the heterocyclic amides to afford the corresponding chloroheterocycles. Secondly, the chloroherocycles and N-cyclohexyl dithiocarbamate cyclohexylammonium salt were heated in chloroform for 12 h at 61 °C to afford heteocyclic thioamides in excellent yields. PMID:28228858

  7. Spindle extraction method for ISAR image based on Radon transform

    NASA Astrophysics Data System (ADS)

    Wei, Xia; Zheng, Sheng; Zeng, Xiangyun; Zhu, Daoyuan; Xu, Gaogui

    2015-12-01

    In this paper, a method of spindle extraction of target in inverse synthetic aperture radar (ISAR) image is proposed which depends on Radon Transform. Firstly, utilizing Radon Transform to detect all straight lines which are collinear with these line segments in image. Then, using Sobel operator to detect image contour. Finally, finding all intersections of each straight line and image contour, the two intersections which have maximum distance between them is the two ends of this line segment and the longest line segment of all line segments is spindle of target. According to the proposed spindle extraction method, one hundred simulated ISAR images which are respectively rotated 0 degrees, 10 degrees, 20 degrees, 30 degrees and 40 degrees in counterclockwise are used to do experiment and the proposed method and the detection results are more close to the real spindle of target than the method based on Hough Transform .

  8. Use of the Transformative Framework in Mixed Methods Studies

    ERIC Educational Resources Information Center

    Sweetman, David; Badiee, Manijeh; Creswell, John W.

    2010-01-01

    A concern exists that mixed methods studies do not contain advocacy stances. Preliminary evidence suggests that this is not the case, but to address this issue in more depth the authors examined 13 mixed methods studies that contained an advocacy, transformative lens. Such a lens consisted of incorporating intent to advocate for an improvement in…

  9. A novel document ranking method using the discrete cosine transform.

    PubMed

    Park, Laurence A F; Palaniswami, Marimuthu; Ramamohanarao, Kotagiri

    2005-01-01

    We propose a new Spectral text retrieval method using the Discrete Cosine Transform (DCT). By taking advantage of the properties of the DCT and by employing the fast query and compression techniques found in vector space methods (VSM), we show that we can process queries as fast as VSM and achieve a much higher precision.

  10. Spatial Fourier transform method for evaluating SQUID gradiometers

    SciTech Connect

    Ribeiro, P.C.; Bruno, A.C.; Paulsen, C.C.; Symko, O.G.

    1987-08-01

    A simple method of measuring the spatial transfer function of a gradiometer, consisting of a flux transformer coupled to a SQUID, is presented and it is compared with theoretical predictions. Based, on this approach, a new method of reporting a gradiometer's performance is proposed; the rejection factor is expressed in decibels obtained directly from the transfer function plot.

  11. Blind digital watermarking method in the fractional Fourier transform domain

    NASA Astrophysics Data System (ADS)

    Lang, Jun; Zhang, Zheng-guang

    2014-02-01

    In this paper, we proposed a novel blind digital image watermarking algorithm based on the fractional Fourier transform (FRFT), which is a generalization of the ordinary Fourier transform and its output has the mixed time and frequency components of the signal. The original image is segmented into non-overlapping blocks for watermarking, and each block is transformed by the two dimensional fractional Fourier transform with two fractional orders. Then each pixel value of binary watermark is embedded by modifying the back-diagonal FRFT coefficients of each image block at the same location with a random array. After perform an inverse two dimensional fractional Fourier transform, we can obtain the watermarked image and the transform orders can be consider as the encryption keys in this method. A series of attacking experiments are performed on the proposed method. The experiments results show that the proposed algorithm not only is of good imperceptibility and security and is very robust to JPEG compression noise attacks and image manipulation operations, but also can provide protection even under compound attacks.

  12. Transforming Mean and Osculating Elements Using Numerical Methods

    NASA Technical Reports Server (NTRS)

    Ely, Todd A.

    2010-01-01

    Mean element propagation of perturbed two body orbits has as its mathematical basis averaging theory of nonlinear dynamical systems. Averaged mean elements define the long-term evolution characteristics of an orbit. Using averaging theory, a near identity transformation can be found that transforms the mean elements back to the osculating elements that contain short period terms in addition to the secular and long period mean elements. The ability to perform the conversion is necessary so that orbit design conducted in mean elements can be converted back into osculating results. In the present work, this near identity transformation is found using the Fast Fourier Transform. An efficient method is found that is capable of recovering the osculating elements to first order

  13. A novel method to transform prokaryotic cells using shock waves

    NASA Astrophysics Data System (ADS)

    Nataraja, K. N.; Udayakumar, M.; Jagadeesh, G.

    The transgenic approach that is being used to study gene function or to improve the efficiency of crop plants/organisms involves transformation of a wide range of cells, tissues, and organisms with nucleic acid. In this study we report a new micro- shock assisted prokaryotic cell transformation technique. An underwater electric discharge based shock wave generator (25 kV; 150 m A; high voltage capacitor) has been designed and fabricated to carry out the prokaryotic cell transformation experiments. Test tubes with bacterial cell suspension with appropriate plasmid DNA, immersed in water are exposed to shock wave loading (typical overpressure 130 bar). The transformation efficiency of samples of the prokaryotic cells exposed to shock waves is very high compared to conventional methods.

  14. [Genetic transformation of buckwheat ( Fagopyrum esculentum Moench ) with AtNHX1 gene and regeneration of salt-tolerant transgenic plants].

    PubMed

    Cheng, Li-Hong; Zhang, Bo; Xu, Zi-Qin

    2007-01-01

    The Arabidopsis thaliana tonoplast Na+ /H+ antiporter gene, AtNHX1, was transferred into buckwheat by Agrobacterium-mediated method. Transgenic buckwheat plants were regenerated and selected on MS basal medium supplemented with 2.0mg/L 6-BA, 1.0mg/L KT, 0.lmg/L IAA, 50mg/L kanamycin and 500mg/L carbenicillin. 426 seedlings from 36 resistant calli originated from 864 explants (transformed about at 4.17 percentage) exhibited resistance to kanamycin. The transformants were confirmed by PCR, Southern blotting, RT-PCR and Northern blotting analysis. After stress treatment for 6 weeks with 200mmol/L NaCl, transgenic plants survived, while wild-type plants did not. After 3 days of stress treatment through different concentrations of NaCl, transgenic plants accumulated higher concentration of Na+ and proline than the control plants. However, the K+ concentration of transgenic plants declined in comparison with the control plants. Moreover, the rutin content of the roots, stems and leaves of transgenic buckwheat increased than those of the control plants. These results showed that it could be possible to improve the salt-tolerance of crops with genetic technology.

  15. Physical methods for genetic transformation of fungi and yeast.

    PubMed

    Rivera, Ana Leonor; Magaña-Ortíz, Denis; Gómez-Lim, Miguel; Fernández, Francisco; Loske, Achim M

    2014-06-01

    The production of transgenic fungi is a routine process. Currently, it is possible to insert genes from other fungi, viruses, bacteria and even animals, albeit with low efficiency, into the genomes of a number of fungal species. Genetic transformation requires the penetration of the transgene through the fungal cell wall, a process that can be facilitated by biological or physical methods. Novel methodologies for the efficient introduction of specific genes and stronger promoters are needed to increase production levels. A possible solution to this problem is the recently discovered shock-wave-mediated transformation. The objective of this article is to review the state of the art of the physical methods used for genetic fungi transformation and to describe some of the basic physics and molecular biology behind them.

  16. Physical methods for genetic transformation of fungi and yeast

    NASA Astrophysics Data System (ADS)

    Rivera, Ana Leonor; Magaña-Ortíz, Denis; Gómez-Lim, Miguel; Fernández, Francisco; Loske, Achim M.

    2014-06-01

    The production of transgenic fungi is a routine process. Currently, it is possible to insert genes from other fungi, viruses, bacteria and even animals, albeit with low efficiency, into the genomes of a number of fungal species. Genetic transformation requires the penetration of the transgene through the fungal cell wall, a process that can be facilitated by biological or physical methods. Novel methodologies for the efficient introduction of specific genes and stronger promoters are needed to increase production levels. A possible solution to this problem is the recently discovered shock-wave-mediated transformation. The objective of this article is to review the state of the art of the physical methods used for genetic fungi transformation and to describe some of the basic physics and molecular biology behind them.

  17. A method to perform a fast fourier transform with primitive image transformations.

    PubMed

    Sheridan, Phil

    2007-05-01

    The Fourier transform is one of the most important transformations in image processing. A major component of this influence comes from the ability to implement it efficiently on a digital computer. This paper describes a new methodology to perform a fast Fourier transform (FFT). This methodology emerges from considerations of the natural physical constraints imposed by image capture devices (camera/eye). The novel aspects of the specific FFT method described include: 1) a bit-wise reversal re-grouping operation of the conventional FFT is replaced by the use of lossless image rotation and scaling and 2) the usual arithmetic operations of complex multiplication are replaced with integer addition. The significance of the FFT presented in this paper is introduced by extending a discrete and finite image algebra, named Spiral Honeycomb Image Algebra (SHIA), to a continuous version, named SHIAC.

  18. Comparison of a discrete wavelet transform method and a modified undecimated discrete wavelet transform method for denoising of mammograms.

    PubMed

    Matsuyama, Eri; Tsai, Du-Yih; Lee, Yongbum; Takahashi, Noriyuki

    2013-01-01

    The purpose of this study was to evaluate the performance of a conventional discrete wavelet transform (DWT) method and a modified undecimated discrete wavelet transform (M-UDWT) method applied to mammographic image denoising. Mutual information, mean square error, and signal to noise ratio were used as image quality measures of images processed by the two methods. We examined the performance of the two methods with visual perceptual evaluation. A two-tailed F test was used to measure statistical significance. The difference between the M-UDWT processed images and the conventional DWT-method processed images was statistically significant (P<0.01). The authors confirmed the superiority and effectiveness of the M-UDWT method. The results of this study suggest the M-UDWT method may provide better image quality as compared to the conventional DWT.

  19. Fourier-transform and global contrast interferometer alignment methods

    DOEpatents

    Goldberg, Kenneth A.

    2001-01-01

    Interferometric methods are presented to facilitate alignment of image-plane components within an interferometer and for the magnified viewing of interferometer masks in situ. Fourier-transforms are performed on intensity patterns that are detected with the interferometer and are used to calculate pseudo-images of the electric field in the image plane of the test optic where the critical alignment of various components is being performed. Fine alignment is aided by the introduction and optimization of a global contrast parameter that is easily calculated from the Fourier-transform.

  20. An efficient method for stable protein targeting in grasses (Poaceae): a case study in Puccinellia tenuiflora

    PubMed Central

    2014-01-01

    Background An efficient transformation method is lacking for most non-model plant species to test gene function. Therefore, subcellular localization of proteins of interest from non-model plants is mainly carried out through transient transformation in homologous cells or in heterologous cells from model species such as Arabidopsis. Although analysis of expression patterns in model organisms like yeast and Arabidopsis can provide important clues about protein localization, these heterologous systems may not always faithfully reflect the native subcellular distribution in other species. On the other hand, transient expression in protoplasts from species of interest has limited ability for detailed sub-cellular localization analysis (e.g., those involving subcellular fractionation or sectioning and immunodetection), as it results in heterogeneous populations comprised of both transformed and untransformed cells. Results We have developed a simple and reliable method for stable transformation of plant cell suspensions that are suitable for protein subcellular localization analyses in the non-model monocotyledonous plant Puccinellia tenuiflora. Optimization of protocols for obtaining suspension-cultured cells followed by Agrobacterium-mediated genetic transformation allowed us to establish stably transformed cell lines, which could be maintained indefinitely in axenic culture supplied with the proper antibiotic. As a case study, protoplasts of transgenic cell lines stably transformed with an ammonium transporter-green fluorescent protein (PutAMT1;1-GFP) fusion were successfully used for subcellular localization analyses in P. tenuiflora. Conclusions We present a reliable method for the generation of stably transformed P. tenuiflora cell lines, which, being available in virtually unlimited amounts, can be conveniently used for any type of protein subcellular localization analysis required. Given its simplicity, the method can be used as reference for other non

  1. A New Shape Description Method Using Angular Radial Transform

    NASA Astrophysics Data System (ADS)

    Lee, Jong-Min; Kim, Whoi-Yul

    Shape is one of the primary low-level image features in content-based image retrieval. In this paper we propose a new shape description method that consists of a rotationally invariant angular radial transform descriptor (IARTD). The IARTD is a feature vector that combines the magnitude and aligned phases of the angular radial transform (ART) coefficients. A phase correction scheme is employed to produce the aligned phase so that the IARTD is invariant to rotation. The distance between two IARTDs is defined by combining differences in the magnitudes and aligned phases. In an experiment using the MPEG-7 shape dataset, the proposed method outperforms existing methods; the average BEP of the proposed method is 57.69%, while the average BEPs of the invariant Zernike moments descriptor and the traditional ART are 41.64% and 36.51%, respectively.

  2. Frequency-dependent FDTD methods using Z transforms

    NASA Technical Reports Server (NTRS)

    Sullivan, Dennis M.

    1992-01-01

    While the frequency-dependent finite-difference time-domain, or (FD)2TD, method can correctly calculate EM propagation through media whose dielectric properties are frequency-dependent, more elaborate applications lead to greater (FD)2TD complexity. Z-transform theory is presently used to develop the mathematical bases of the (FD)2TD method, simultaneously obtaining a clearer formulation and allowing researchers to draw on the existing literature of systems analysis and signal-processing.

  3. A dynamic integrated fault diagnosis method for power transformers.

    PubMed

    Gao, Wensheng; Bai, Cuifen; Liu, Tong

    2015-01-01

    In order to diagnose transformer fault efficiently and accurately, a dynamic integrated fault diagnosis method based on Bayesian network is proposed in this paper. First, an integrated fault diagnosis model is established based on the causal relationship among abnormal working conditions, failure modes, and failure symptoms of transformers, aimed at obtaining the most possible failure mode. And then considering the evidence input into the diagnosis model is gradually acquired and the fault diagnosis process in reality is multistep, a dynamic fault diagnosis mechanism is proposed based on the integrated fault diagnosis model. Different from the existing one-step diagnosis mechanism, it includes a multistep evidence-selection process, which gives the most effective diagnostic test to be performed in next step. Therefore, it can reduce unnecessary diagnostic tests and improve the accuracy and efficiency of diagnosis. Finally, the dynamic integrated fault diagnosis method is applied to actual cases, and the validity of this method is verified.

  4. A Dynamic Integrated Fault Diagnosis Method for Power Transformers

    PubMed Central

    Gao, Wensheng; Liu, Tong

    2015-01-01

    In order to diagnose transformer fault efficiently and accurately, a dynamic integrated fault diagnosis method based on Bayesian network is proposed in this paper. First, an integrated fault diagnosis model is established based on the causal relationship among abnormal working conditions, failure modes, and failure symptoms of transformers, aimed at obtaining the most possible failure mode. And then considering the evidence input into the diagnosis model is gradually acquired and the fault diagnosis process in reality is multistep, a dynamic fault diagnosis mechanism is proposed based on the integrated fault diagnosis model. Different from the existing one-step diagnosis mechanism, it includes a multistep evidence-selection process, which gives the most effective diagnostic test to be performed in next step. Therefore, it can reduce unnecessary diagnostic tests and improve the accuracy and efficiency of diagnosis. Finally, the dynamic integrated fault diagnosis method is applied to actual cases, and the validity of this method is verified. PMID:25685841

  5. A transformation method for constrained-function minimization

    NASA Technical Reports Server (NTRS)

    Park, S. K.

    1975-01-01

    A direct method for constrained-function minimization is discussed. The method involves the construction of an appropriate function mapping all of one finite dimensional space onto the region defined by the constraints. Functions which produce such a transformation are constructed for a variety of constraint regions including, for example, those arising from linear and quadratic inequalities and equalities. In addition, the computational performance of this method is studied in the situation where the Davidon-Fletcher-Powell algorithm is used to solve the resulting unconstrained problem. Good performance is demonstrated for 19 test problems by achieving rapid convergence to a solution from several widely separated starting points.

  6. A method for decoding the neurophysiological spike-response transform.

    PubMed

    Stern, Estee; García-Crescioni, Keyla; Miller, Mark W; Peskin, Charles S; Brezina, Vladimir

    2009-11-15

    Many physiological responses elicited by neuronal spikes-intracellular calcium transients, synaptic potentials, muscle contractions-are built up of discrete, elementary responses to each spike. However, the spikes occur in trains of arbitrary temporal complexity, and each elementary response not only sums with previous ones, but can itself be modified by the previous history of the activity. A basic goal in system identification is to characterize the spike-response transform in terms of a small number of functions-the elementary response kernel and additional kernels or functions that describe the dependence on previous history-that will predict the response to any arbitrary spike train. Here we do this by developing further and generalizing the "synaptic decoding" approach of Sen et al. (1996). Given the spike times in a train and the observed overall response, we use least-squares minimization to construct the best estimated response and at the same time best estimates of the elementary response kernel and the other functions that characterize the spike-response transform. We avoid the need for any specific initial assumptions about these functions by using techniques of mathematical analysis and linear algebra that allow us to solve simultaneously for all of the numerical function values treated as independent parameters. The functions are such that they may be interpreted mechanistically. We examine the performance of the method as applied to synthetic data. We then use the method to decode real synaptic and muscle contraction transforms.

  7. Recent advances in soybean transformation and their application to molecular breeding and genomic analysis

    PubMed Central

    Yamada, Tetsuya; Takagi, Kyoko; Ishimoto, Masao

    2012-01-01

    Herbicide-resistant transgenic soybean plants hold a leading market share in the USA and other countries, but soybean has been regarded as recalcitrant to transformation for many years. The cumulative and, at times, exponential advances in genetic manipulation have made possible further choices for soybean transformation. The most widely and routinely used transformation systems are cotyledonary node–Agrobacterium-mediated transformation and somatic embryo–particle-bombardment-mediated transformation. These ready systems enable us to improve seed qualities and agronomic characteristics by transgenic approaches. In addition, with the accumulation of soybean genomic resources, convenient or promising approaches will be requisite for the determination and use of gene function in soybean. In this article, we describe recent advances in and problems of soybean transformation, and survey the current transgenic approaches for applied and basic research in Japan. PMID:23136488

  8. In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance.

    PubMed

    Pandey, Sonika; Patel, Manish Kumar; Mishra, Avinash; Jha, Bhavanath

    2016-01-01

    Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas.

  9. In planta Transformed Cumin (Cuminum cyminum L.) Plants, Overexpressing the SbNHX1 Gene Showed Enhanced Salt Endurance

    PubMed Central

    Pandey, Sonika; Patel, Manish Kumar; Jha, Bhavanath

    2016-01-01

    Cumin is an annual, herbaceous, medicinal, aromatic, spice glycophyte that contains diverse applications as a food and flavoring additive, and therapeutic agents. An efficient, less time consuming, Agrobacterium-mediated, a tissue culture-independent in planta genetic transformation method was established for the first time using cumin seeds. The SbNHX1 gene, cloned from an extreme halophyte Salicornia brachiata was transformed in cumin using optimized in planta transformation method. The SbNHX1 gene encodes a vacuolar Na+/H+ antiporter and is involved in the compartmentalization of excess Na+ ions into the vacuole and maintenance of ion homeostasis Transgenic cumin plants were confirmed by PCR using gene (SbNHX1, uidA and hptII) specific primers. The single gene integration event and overexpression of the gene were confirmed by Southern hybridization and competitive RT-PCR, respectively. Transgenic lines L3 and L13 showed high expression of the SbNHX1 gene compared to L6 whereas moderate expression was detected in L5 and L10 transgenic lines. Transgenic lines (L3, L5, L10 and L13), overexpressing the SbNHX1 gene, showed higher photosynthetic pigments (chlorophyll a, b and carotenoid), and lower electrolytic leakage, lipid peroxidation (MDA content) and proline content as compared to wild type plants under salinity stress. Though transgenic lines were also affected by salinity stress but performed better compared to WT plants. The ectopic expression of the SbNHX1 gene confirmed enhanced salinity stress tolerance in cumin as compared to wild type plants under stress condition. The present study is the first report of engineering salt tolerance in cumin, so far and the plant may be utilized for the cultivation in saline areas. PMID:27411057

  10. Electrocardiogram ST-Segment Morphology Delineation Method Using Orthogonal Transformations

    PubMed Central

    2016-01-01

    Differentiation between ischaemic and non-ischaemic transient ST segment events of long term ambulatory electrocardiograms is a persisting weakness in present ischaemia detection systems. Traditional ST segment level measuring is not a sufficiently precise technique due to the single point of measurement and severe noise which is often present. We developed a robust noise resistant orthogonal-transformation based delineation method, which allows tracing the shape of transient ST segment morphology changes from the entire ST segment in terms of diagnostic and morphologic feature-vector time series, and also allows further analysis. For these purposes, we developed a new Legendre Polynomials based Transformation (LPT) of ST segment. Its basis functions have similar shapes to typical transient changes of ST segment morphology categories during myocardial ischaemia (level, slope and scooping), thus providing direct insight into the types of time domain morphology changes through the LPT feature-vector space. We also generated new Karhunen and Lo ève Transformation (KLT) ST segment basis functions using a robust covariance matrix constructed from the ST segment pattern vectors derived from the Long Term ST Database (LTST DB). As for the delineation of significant transient ischaemic and non-ischaemic ST segment episodes, we present a study on the representation of transient ST segment morphology categories, and an evaluation study on the classification power of the KLT- and LPT-based feature vectors to classify between ischaemic and non-ischaemic ST segment episodes of the LTST DB. Classification accuracy using the KLT and LPT feature vectors was 90% and 82%, respectively, when using the k-Nearest Neighbors (k = 3) classifier and 10-fold cross-validation. New sets of feature-vector time series for both transformations were derived for the records of the LTST DB which is freely available on the PhysioNet website and were contributed to the LTST DB. The KLT and LPT

  11. Method to transform algae, materials therefor, and products produced thereby

    DOEpatents

    Dunahay, T.G.; Roessler, P.G.; Jarvis, E.E.

    1997-08-26

    Disclosed is a method to transform chlorophyll C-containing algae. The method includes introducing a recombinant molecule comprising a nucleic acid molecule encoding a dominant selectable marker operatively linked to an algal regulatory control sequence into a chlorophyll C-containing alga in such a manner that the marker is produced by the alga. In a preferred embodiment the algal regulatory control sequence is derived from a diatom and preferably Cyclotella cryptica. Also disclosed is a chimeric molecule having one or more regulatory control sequences derived from one or more chlorophyll C-containing algae operatively linked to a nucleic acid molecule encoding a selectable marker, an RNA molecule and/or a protein, wherein the nucleic acid molecule does not normally occur with one or more of the regulatory control sequences. Further, specifically disclosed are molecules pACCNPT10, pACCNPT4.8 and pACCNPT5.1. The methods and materials of the present invention provide the ability to accomplish stable genetic transformation of chlorophyll C-containing algae. 2 figs.

  12. Transformation and Evaluation of Cry1Ac+Cry2A and GTGene in Gossypium hirsutum L.

    PubMed Central

    Puspito, Agung N.; Rao, Abdul Q.; Hafeez, Muhammad N.; Iqbal, Muhammad S.; Bajwa, Kamran S.; Ali, Qurban; Rashid, Bushra; Abbas, Muhammad A.; Latif, Ayesha; Shahid, Ahmad A.; Nasir, Idrees A.; Husnain, Tayyab

    2015-01-01

    More than 50 countries around the globe cultivate cotton on a large scale. It is a major cash crop of Pakistan and is considered “white gold” because it is highly important to the economy of Pakistan. In addition to its importance, cotton cultivation faces several problems, such as insect pests, weeds, and viruses. In the past, insects have been controlled by insecticides, but this method caused a severe loss to the economy. However, conventional breeding methods have provided considerable breakthroughs in the improvement of cotton, but it also has several limitations. In comparison with conventional methods, biotechnology has the potential to create genetically modified plants that are environmentally safe and economically viable. In this study, a local cotton variety VH 289 was transformed with two Bt genes (Cry1Ac and Cry2A) and a herbicide resistant gene (cp4 EPSPS) using the Agrobacterium mediated transformation method. The constitutive CaMV 35S promoter was attached to the genes taken from Bacillus thuringiensis (Bt) and to an herbicide resistant gene during cloning, and this promoter was used for the expression of the genes in cotton plants. This construct was used to develop the Glyphosate Tolerance Gene (GTGene) for herbicide tolerance and insecticidal gene (Cry1Ac and Cry2A) for insect tolerance in the cotton variety VH 289. The transgenic cotton variety performed 85% better compared with the non-transgenic variety. The study results suggest that farmers should use the transgenic cotton variety for general cultivation to improve the production of cotton. PMID:26617613

  13. Enhanced salt tolerance of alfalfa (Medicago sativa) by rstB gene transformation.

    PubMed

    Zhang, Wan-Jun; Wang, Tao

    2015-05-01

    Generating salt tolerance forage plant is essential for use of the land affected by high salinity. A salt tolerance gene rstB was used as a selectable marker gene in Agrobacterium-mediated transformation of tobacco under a selective regime of 170mM NaCl. The transgenic plants showed clear improvement in salt tolerance. To improve salt tolerance of alfalfa (Medicago sativa L.), rstB gene was introduced into alfalfa genome by Agrobacterium-mediated transformation. No abnormal phenotype was observed among the transgenic plants when compared with wild type (wt) plants. Significant enhancement of resistance to salt-shock treatment was noted on the rstB transgenic (T0) plants. Transgenic second-generation (T1) seeds showed improved germination rate and seedling growth under salt-stress condition. Hindered Na(+) accumulation, but enhanced Ca(2+) accumulation was observed on the rstB T1 plants when subjected to salt-stresses. Enhanced calcium accumulation in transgenic plants was also verified by cytohistochemical localization of calcium. Under salt-stress of 50mM NaCl, about 15% of the transgenic plants finished their life-cycle but the wt plants had no flower formation. The results demonstrated that the expression of rstB gene improved salt tolerance in transgenic alfalfa.

  14. Transformational Teaching: Theoretical Underpinnings, Basic Principles, and Core Methods

    PubMed Central

    Slavich, George M.; Zimbardo, Philip G.

    2012-01-01

    Approaches to classroom instruction have evolved considerably over the past 50 years. This progress has been spurred by the development of several learning principles and methods of instruction, including active learning, student-centered learning, collaborative learning, experiential learning, and problem-based learning. In the present paper, we suggest that these seemingly different strategies share important underlying characteristics and can be viewed as complimentary components of a broader approach to classroom instruction called transformational teaching. Transformational teaching involves creating dynamic relationships between teachers, students, and a shared body of knowledge to promote student learning and personal growth. From this perspective, instructors are intellectual coaches who create teams of students who collaborate with each other and with their teacher to master bodies of information. Teachers assume the traditional role of facilitating students’ acquisition of key course concepts, but do so while enhancing students’ personal development and attitudes toward learning. They accomplish these goals by establishing a shared vision for a course, providing modeling and mastery experiences, challenging and encouraging students, personalizing attention and feedback, creating experiential lessons that transcend the boundaries of the classroom, and promoting ample opportunities for preflection and reflection. We propose that these methods are synergistically related and, when used together, maximize students’ potential for intellectual and personal growth. PMID:23162369

  15. Method to transform algae, materials therefor, and products produced thereby

    DOEpatents

    Dunahay, Terri Goodman; Roessler, Paul G.; Jarvis, Eric E.

    1997-01-01

    Disclosed is a method to transform chlorophyll C-containing algae which includes introducing a recombinant molecule comprising a nucleic acid molecule encoding a dominant selectable marker operatively linked to an algal regulatory control sequence into a chlorophyll C-containing alga in such a manner that the marker is produced by the alga. In a preferred embodiment the algal regulatory control sequence is derived from a diatom and preferably Cyclotella cryptica. Also disclosed is a chimeric molecule having one or more regulatory control sequences derived from one or more chlorophyll C-containing algae operatively linked to a nucleic acid molecule encoding a selectable marker, an RNA molecule and/or a protein, wherein the nucleic acid molecule does not normally occur with one or more of the regulatory control sequences. Further specifically disclosed are molecules pACCNPT10, pACCNPT4.8 and pACCNPT5.1. The methods and materials of the present invention provide the ability to accomplish stable genetic transformation of chlorophyll C-containing algae.

  16. Transformational Teaching: Theoretical Underpinnings, Basic Principles, and Core Methods.

    PubMed

    Slavich, George M; Zimbardo, Philip G

    2012-12-01

    Approaches to classroom instruction have evolved considerably over the past 50 years. This progress has been spurred by the development of several learning principles and methods of instruction, including active learning, student-centered learning, collaborative learning, experiential learning, and problem-based learning. In the present paper, we suggest that these seemingly different strategies share important underlying characteristics and can be viewed as complimentary components of a broader approach to classroom instruction called transformational teaching. Transformational teaching involves creating dynamic relationships between teachers, students, and a shared body of knowledge to promote student learning and personal growth. From this perspective, instructors are intellectual coaches who create teams of students who collaborate with each other and with their teacher to master bodies of information. Teachers assume the traditional role of facilitating students' acquisition of key course concepts, but do so while enhancing students' personal development and attitudes toward learning. They accomplish these goals by establishing a shared vision for a course, providing modeling and mastery experiences, challenging and encouraging students, personalizing attention and feedback, creating experiential lessons that transcend the boundaries of the classroom, and promoting ample opportunities for preflection and reflection. We propose that these methods are synergistically related and, when used together, maximize students' potential for intellectual and personal growth.

  17. Combined chemical and physical transformation method with RbCl and sepiolite for the transformation of various bacterial species.

    PubMed

    Ren, Jun; Lee, Haram; Yoo, Seung Min; Yu, Myeong-Sang; Park, Hansoo; Na, Dokyun

    2017-04-01

    DNA transformation that delivers plasmid DNAs into bacterial cells is fundamental in genetic manipulation to engineer and study bacteria. Developed transformation methods to date are optimized to specific bacterial species for high efficiency. Thus, there is always a demand for simple and species-independent transformation methods. We herein describe the development of a chemico-physical transformation method that combines a rubidium chloride (RbCl)-based chemical method and sepiolite-based physical method, and report its use for the simple and efficient delivery of DNA into various bacterial species. Using this method, the best transformation efficiency for Escherichia coli DH5α was 4.3×10(6)CFU/μg of pUC19 plasmid, which is higher than or comparable to the reported transformation efficiencies to date. This method also allowed the introduction of plasmid DNAs into Bacillus subtilis (5.7×10(3)CFU/μg of pSEVA3b67Rb), Bacillus megaterium (2.5×10(3)CFU/μg of pSPAsp-hp), Lactococcus lactis subsp. lactis (1.0×10(2)CFU/μg of pTRKH3-ermGFP), and Lactococcus lactis subsp. cremoris (2.2×10(2)CFU/μg of pMSP3535VA). Remarkably, even when the conventional chemical and physical methods failed to generate transformed cells in Bacillus sp. and Enterococcus faecalis, E. malodoratus and E. mundtii, our combined method showed a significant transformation efficiency (2.4×10(4), 4.5×10(2), 2×10(1), and 0.5×10(1)CFU/μg of plasmid DNA). Based on our results, we anticipate that our simple and efficient transformation method should prove usefulness for introducing DNA into various bacterial species without complicated optimization of parameters affecting DNA entry into the cell.

  18. TRANSFORMATION

    SciTech Connect

    LACKS,S.A.

    2003-10-09

    Transformation, which alters the genetic makeup of an individual, is a concept that intrigues the human imagination. In Streptococcus pneumoniae such transformation was first demonstrated. Perhaps our fascination with genetics derived from our ancestors observing their own progeny, with its retention and assortment of parental traits, but such interest must have been accelerated after the dawn of agriculture. It was in pea plants that Gregor Mendel in the late 1800s examined inherited traits and found them to be determined by physical elements, or genes, passed from parents to progeny. In our day, the material basis of these genetic determinants was revealed to be DNA by the lowly bacteria, in particular, the pneumococcus. For this species, transformation by free DNA is a sexual process that enables cells to sport new combinations of genes and traits. Genetic transformation of the type found in S. pneumoniae occurs naturally in many species of bacteria (70), but, initially only a few other transformable species were found, namely, Haemophilus influenzae, Neisseria meningitides, Neisseria gonorrheae, and Bacillus subtilis (96). Natural transformation, which requires a set of genes evolved for the purpose, contrasts with artificial transformation, which is accomplished by shocking cells either electrically, as in electroporation, or by ionic and temperature shifts. Although such artificial treatments can introduce very small amounts of DNA into virtually any type of cell, the amounts introduced by natural transformation are a million-fold greater, and S. pneumoniae can take up as much as 10% of its cellular DNA content (40).

  19. Spectrum-transformed sequential testing method for signal validation applications

    SciTech Connect

    Gross, K.C.; Hoyer, K.K.

    1992-06-01

    The Sequential Probability Ratio Test (SPRT) has proven to be a valuable tool in a variety of reactor applications for signal validation and for sensor and equipment operability surveillance. One drawback of the conventional SPRT method is that its domain of application is limited to signals that are contaminated by gaussian white noise. Nongaussian process variables contaminated by serial correlation can produce higher-than-specified rates of false alarms and missed alarms for SPRT-based surveillance systems. To overcome this difficulty we present here the development and computer implementation of a new technique, the spectrum-transformed sequential testing method. This method retains the excellent surveillance advantage of the SPRT (extremely high sensitivity for very early annunciation of the onset of disturbances in monitored signals), and its false-alarm and missed-alarm probabilities are unaffected by the presence of serial correlation in the data. Example applications of the new method to serially-correlated reactor variables are demonstrated using data recorded from EBR-II.

  20. Spectrum-transformed sequential testing method for signal validation applications

    SciTech Connect

    Gross, K.C. ); Hoyer, K.K. . Dept. of Industrial Engineering and Management Sciences)

    1992-01-01

    The Sequential Probability Ratio Test (SPRT) has proven to be a valuable tool in a variety of reactor applications for signal validation and for sensor and equipment operability surveillance. One drawback of the conventional SPRT method is that its domain of application is limited to signals that are contaminated by gaussian white noise. Nongaussian process variables contaminated by serial correlation can produce higher-than-specified rates of false alarms and missed alarms for SPRT-based surveillance systems. To overcome this difficulty we present here the development and computer implementation of a new technique, the spectrum-transformed sequential testing method. This method retains the excellent surveillance advantage of the SPRT (extremely high sensitivity for very early annunciation of the onset of disturbances in monitored signals), and its false-alarm and missed-alarm probabilities are unaffected by the presence of serial correlation in the data. Example applications of the new method to serially-correlated reactor variables are demonstrated using data recorded from EBR-II.

  1. Method and apparatus for calibrating a linear variable differential transformer

    DOEpatents

    Pokrywka, Robert J.

    2005-01-18

    A calibration apparatus for calibrating a linear variable differential transformer (LVDT) having an armature positioned in au LVDT armature orifice, and the armature able to move along an axis of movement. The calibration apparatus includes a heating mechanism with an internal chamber, a temperature measuring mechanism for measuring the temperature of the LVDT, a fixture mechanism with an internal chamber for at least partially accepting the LVDT and for securing the LVDT within the heating mechanism internal chamber, a moving mechanism for moving the armature, a position measurement mechanism for measuring the position of the armature, and an output voltage measurement mechanism. A method for calibrating an LVDT, including the steps of: powering the LVDT; heating the LVDT to a desired temperature; measuring the position of the armature with respect to the armature orifice; and measuring the output voltage of the LVDT.

  2. TRANSFORMER

    DOEpatents

    Baker, W.R.

    1959-08-25

    Transformers of a type adapted for use with extreme high power vacuum tubes where current requirements may be of the order of 2,000 to 200,000 amperes are described. The transformer casing has the form of a re-entrant section being extended through an opening in one end of the cylinder to form a coaxial terminal arrangement. A toroidal multi-turn primary winding is disposed within the casing in coaxial relationship therein. In a second embodiment, means are provided for forming the casing as a multi-turn secondary. The transformer is characterized by minimized resistance heating, minimized external magnetic flux, and an economical construction.

  3. 3-D surface profilometry based on modulation measurement by applying wavelet transform method

    NASA Astrophysics Data System (ADS)

    Zhong, Min; Chen, Feng; Xiao, Chao; Wei, Yongchao

    2017-01-01

    A new analysis of 3-D surface profilometry based on modulation measurement technique by the application of Wavelet Transform method is proposed. As a tool excelling for its multi-resolution and localization in the time and frequency domains, Wavelet Transform method with good localized time-frequency analysis ability and effective de-noizing capacity can extract the modulation distribution more accurately than Fourier Transform method. Especially for the analysis of complex object, more details of the measured object can be well remained. In this paper, the theoretical derivation of Wavelet Transform method that obtains the modulation values from a captured fringe pattern is given. Both computer simulation and elementary experiment are used to show the validity of the proposed method by making a comparison with the results of Fourier Transform method. The results show that the Wavelet Transform method has a better performance than the Fourier Transform method in modulation values retrieval.

  4. On optimal methods for adiabatic quantum state transformations

    NASA Astrophysics Data System (ADS)

    Somma, Rolando

    2013-03-01

    Many problems in science could be solved by preparing the low-energy quantum state (or any eigenstate) of a Hamiltonian. A common example is the Boolean satisfiability problem, where each clause can be mapped to the energy of an interacting many-body system, and the problem reduces to minimizing the energy. In quantum computing, adiabatic quantum state transformations (ASTs) provide a tool for preparing the quantum state. ASTs are conventionally implemented via slow or adiabatic perturbations to the Hamiltonian, relying on the quantum adiabatic theorem. Nevertheless, more efficient implementations of ASTs exist. In this talk I will review recently developed methods for ASTs that are more efficient and require less assumptions on the Hamiltonians than the conventional implementation. Such methods involve measurements of the states along the evolution path and have a best-case implementation cost of L/G, where L is the length of the (evolved) state path and G is a lower bound to the spectral gap of the Hamiltonians. I will show that this cost is optimal and comment on results of the gap amplification problem, where the goal is to reduce the cost by increasing G. We acknowledge support from NSF through the CCF program and the LDRD programs at Los Alamos National Laboratory and Sandia National Laboratories.

  5. Method of Estimating Continuous Cooling Transformation Curves of Glasses

    NASA Technical Reports Server (NTRS)

    Zhu, Dongmei; Zhou, Wancheng; Ray, Chandra S.; Day, Delbert E.

    2006-01-01

    A method is proposed for estimating the critical cooling rate and continuous cooling transformation (CCT) curve from isothermal TTT data of glasses. The critical cooling rates and CCT curves for a group of lithium disilicate glasses containing different amounts of Pt as nucleating agent estimated through this method are compared with the experimentally measured values. By analysis of the experimental and calculated data of the lithium disilicate glasses, a simple relationship between the crystallized amount in the glasses during continuous cooling, X, and the temperature of undercooling, (Delta)T, was found to be X = AR(sup-4)exp(B (Delta)T), where (Delta)T is the temperature difference between the theoretical melting point of the glass composition and the temperature in discussion, R is the cooling rate, and A and B are constants. The relation between the amount of crystallisation during continuous cooling and isothermal hold can be expressed as (X(sub cT)/X(sub iT) = (4/B)(sup 4) (Delta)T(sup -4), where X(sub cT) stands for the crystallised amount in a glass during continuous cooling for a time t when the temperature comes to T, and X(sub iT) is the crystallised amount during isothermal hold at temperature T for a time t.

  6. Co-transforming bar and CsALDH Genes Enhanced Resistance to Herbicide and Drought and Salt Stress in Transgenic Alfalfa (Medicago sativa L.)

    PubMed Central

    Duan, Zhen; Zhang, Daiyu; Zhang, Jianquan; Di, Hongyan; Wu, Fan; Hu, Xiaowen; Meng, Xuanchen; Luo, Kai; Zhang, Jiyu; Wang, Yanrong

    2015-01-01

    Drought and high salinity are two major abiotic factors that restrict the productivity of alfalfa. By application of the Agrobacterium-mediated transformation method, an oxidative responsive gene, CsALDH12A1, from the desert grass Cleistogenes songorica together with the bar gene associated with herbicide resistance, were co-transformed into alfalfa (Medicago sativa L.). From the all 90 transformants, 16 were positive as screened by spraying 1 mL L-1 10% Basta solution and molecularly diagnosis using PCR. Real-time PCR analysis indicated that drought and salt stress induced high CsALDH expression in the leaves of the transgenic plants. The CsALDH expression levels under drought (15 d) and salt stress (200 mM NaCl) were 6.11 and 6.87 times higher than in the control plants, respectively. In comparison to the WT plants, no abnormal phenotypes were observed among the transgenic plants, which showed significant enhancement of tolerance to 15 d of drought and 10 d of salinity treatment. Evaluation of the physiological and biochemical indices during drought and salt stress of the transgenic plants revealed relatively lower Na+ content and higher K+ content in the leaves relative to the WT plants, a reduction of toxic on effects and maintenance of osmotic adjustment. In addition, the transgenic plants could maintain a higher relative water content level, higher shoot biomass, fewer changes in the photosystem, decreased membrane injury, and a lower level of osmotic stress. These results indicate that the co-expression of the introduced bar and CsALDH genes enhanced the herbicide, drought and salt tolerance of alfalfa and therefore can potentially be used as a novel genetic resource for the future breeding programs to develop new cultivars. PMID:26734025

  7. Integral Transform Methods: A Critical Review of Various Kernels

    NASA Astrophysics Data System (ADS)

    Orlandini, Giuseppina; Turro, Francesco

    2017-03-01

    Some general remarks about integral transform approaches to response functions are made. Their advantage for calculating cross sections at energies in the continuum is stressed. In particular we discuss the class of kernels that allow calculations of the transform by matrix diagonalization. A particular set of such kernels, namely the wavelets, is tested in a model study.

  8. Category's analysis and operational project capacity method of transformation in design

    NASA Astrophysics Data System (ADS)

    Obednina, S. V.; Bystrova, T. Y.

    2015-10-01

    The method of transformation is attracting widespread interest in fields such contemporary design. However, in theory of design little attention has been paid to a categorical status of the term "transformation". This paper presents the conceptual analysis of transformation based on the theory of form employed in the influential essays by Aristotle and Thomas Aquinas. In the present work the transformation as a method of shaping design has been explored as well as potential application of this term in design has been demonstrated.

  9. Genetic transformation of Begonia tuberhybrida by Ri rol genes.

    PubMed

    Kiyokawa, S; Kikuchi, Y; Kamada, H; Harada, H

    1996-04-01

    We have developed an Agrobacterium -mediated transformation system for commercial Begonia species. The leaf explants of Begonia semperflorens, Begonia x hiemalis and B. tuberhybrida were inoculated with Agrobacterium tumefaciens LBA4404 harboring a binary vector pBI121 which contains rolA, B and C genes of an agropine type Ri plasmid (pRiA4b). Kanamycin resistant shoots of B. tuberhybrida were obtained on MS agar medium supplemented with 0.1 mg/l NAA, 0.5 mg/l BA, 500 mg/l claforan and 100 mg/l kanamycin. These shoots exhibited GUS activity and Southern analysis showed a single copy insertion into the genome. When the transgenic plants were transferred to soil, they displayed the phenotype specific to the transgenic plants by A. rhizogenes such as dwarfness, delay of flowering, and wrinkled leaves and petals.

  10. Highly efficient transformation system for Malassezia furfur and Malassezia pachydermatis using Agrobacterium tumefaciens-mediated transformation.

    PubMed

    Celis, A M; Vos, A M; Triana, S; Medina, C A; Escobar, N; Restrepo, S; Wösten, H A B; de Cock, H

    2017-03-01

    Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M. furfur and M. pachydermatis by combining the transformation protocols of Agaricus bisporus and Cryptococcus neoformans. Optimal temperature and co-cultivation time for transformation were 5 and 7days at 19°C and 24°C, respectively. Transformation efficiency was 0.75-1.5% for M. furfur and 0.6-7.5% for M. pachydermatis. Integration of the hpt resistance cassette and gfp was verified using PCR and fluorescence microscopy, respectively. The T-DNA was mitotically stable in approximately 80% of the transformants after 10 times sub-culturing in the absence of hygromycin. Improving transformation protocols contribute to study the biology and pathophysiology of Malassezia.

  11. Transformation Methods for Using Combination of Remotely Sensed Data and Cadastral Maps

    NASA Astrophysics Data System (ADS)

    Dönmez, Ş. Ö.; Tunc, A.

    2016-06-01

    In order to examine using cadastral maps as base maps for aerial orthophotos, two different 2D transformation methods were applied between various coordinate systems. Study area was chosen from Kagithane district in Istanbul. The used data is an orthophoto (30 cm spatial resolution), and cadastral map (1:1000) taken from land office, containing the same region. Transformation methods are chosen as; 1st Order Polynomial Transformation and Helmert 2D Transformation within this study. The test points, used to determine the coefficients between the datums, were 26 common traverse points and the check points, used to compare the transformed coordinates to reliable true coordinates, were 10 common block corners. The transformation methods were applied using Matlab software. After applying the methods, residuals were calculated and compared between each transformation method in order to use cadastral maps as reliable vector data.

  12. Methods for performing fast discrete curvelet transforms of data

    DOEpatents

    Candes, Emmanuel; Donoho, David; Demanet, Laurent

    2010-11-23

    Fast digital implementations of the second generation curvelet transform for use in data processing are disclosed. One such digital transformation is based on unequally-spaced fast Fourier transforms (USFFT) while another is based on the wrapping of specially selected Fourier samples. Both digital transformations return a table of digital curvelet coefficients indexed by a scale parameter, an orientation parameter, and a spatial location parameter. Both implementations are fast in the sense that they run in about O(n.sup.2 log n) flops for n by n Cartesian arrays or about O(N log N) flops for Cartesian arrays of size N=n.sup.3; in addition, they are also invertible, with rapid inversion algorithms of about the same complexity.

  13. Computationally efficient method for Fourier transform of highly chirped pulses for laser and parametric amplifier modeling.

    PubMed

    Andrianov, Alexey; Szabo, Aron; Sergeev, Alexander; Kim, Arkady; Chvykov, Vladimir; Kalashnikov, Mikhail

    2016-11-14

    We developed an improved approach to calculate the Fourier transform of signals with arbitrary large quadratic phase which can be efficiently implemented in numerical simulations utilizing Fast Fourier transform. The proposed algorithm significantly reduces the computational cost of Fourier transform of a highly chirped and stretched pulse by splitting it into two separate transforms of almost transform limited pulses, thereby reducing the required grid size roughly by a factor of the pulse stretching. The application of our improved Fourier transform algorithm in the split-step method for numerical modeling of CPA and OPCPA shows excellent agreement with standard algorithms.

  14. [Construction of the plant expression vector with hepatitis a capsid protein fusion gene and genetic transformation of Citrus. Sinensis Osbeck].

    PubMed

    Hu, Rong; Wei, Hong; Chen, Shan-Chun; He, Yong-Rui

    2004-07-01

    The use of edible plants for the production and delivery of vaccine proteins could provide an economical alternative to fermentation systems. The construction of the plant expression vector pBI121-A was reported, which contained a fusion gene encoding hepatitis A capsid proteins. The gene was located between the left and right Ti border sequences under the control of CaMV35S promoter. The vector was identified via PCR and restriction enzyme analysis and was introduced into Agrobacterium tumerifacience LBA4404. The transgenic Citrus plants were produced by Agrobacterium-mediated transformation of epicotyl segments.13 putatively transformed plants through the kanamycin selection were micrografted onto the seedlings. The presence and integration of the transgene had been verified by PCR analysis. The result showed that five transformants were integrated and the transformation efficiency was 4.1%.

  15. Strain analysis by mismatch moire method and grid method using Fourier transform

    NASA Astrophysics Data System (ADS)

    Morimoto, Y.; Seguchi, Y.; Higashi, T.

    1990-01-01

    We have formerly presented a new method of the moire analysis of strain using the Fourier transform. It uses the phase information of the moire fringe brightness. By shifting the Fourier spectrum of the image of deformed grating lines, we obtain the “complex moire pattern”. Strain distribution is given as the derivatives of the phases of the complex moire fringes. The analysis is completely automated by digital image processing. All of the laborious and subjective procedures required in the conventional analysis such as fringe sign determination, fringe ordering and fringe interpolation are thus eliminated, and objective, fast and accurate analysis can be made. In this paper, we develop the method to a mismatch method and a grid method. We show some applications for analyzing strain distribution by using this method.

  16. Genetic transformation and expression of transgenic lines of Populus x euramericana with insect-resistance and salt-tolerance genes.

    PubMed

    Yang, R L; Wang, A X; Zhang, J; Dong, Y; Yang, M S; Wang, J M

    2016-04-29

    We characterized new transgenic varieties of poplar with multiple insect-resistant and salt stress tolerant genes. Two insect-resistant Bacillus thuringiensis (Bt) genes, Cry1Ac and Cry3A, and a salt-tolerant gene, Betaine aldehyde dehydrogenase (BADH) were inserted into a vector, p209-Cry1Ac-Cry3A-BADH. The clone of Populus x euramericana was transformed by the vector using the Agrobacterium-mediated method. Three transgenic lines were assessed using genetic detection and resistance expression analysis. PCR revealed that exogenous genes Cry1Ac, Cry3A, BADH and selective marker gene NPTII were present in three transgenic lines. Quantitative real-time PCR (qPCR) showed significant differences in the transcriptional abundance of three exogenous genes in different lines. Results of assays for Bt toxic proteins showed that the Cry1Ac and Cry3A toxic protein content of each line was 12.83-26.32 and 2108.91-2724.79 ng/g, respectively. The Cry1Ac toxic protein content of different lines was significantly different; the Cry3A toxic protein content was about 100 times higher than that of the Cry1Ac toxic protein. The insect-resistance test revealed the mortality rate of transgenic lines to Hyphantria cunea L1 larvae varied by 42.2-66.7%, which was significantly higher than non-transgenic lines. The mortality rate of L1 and L2 Plagiodera versicolora larvae was 100%. The insecticidal effect of transgenic lines to P. versicolora larvae was higher than that to H. cunea larvae. NaCl stress tolerance of three transgenic lines under 3-6% NaCl concentration was significantly higher than that of non-transgenic lines.

  17. Analysis of temperature rise for piezoelectric transformer using finite-element method.

    PubMed

    Joo, Hyun-Woo; Lee, Chang-Hwan; Rho, Jong-Seok; Jung, Hyun-Kyo

    2006-08-01

    Analysis of heat problem and temperature field of a piezoelectric transformer, operated at steady-state conditions, is described. The resonance frequency of the transformer is calculated from impedance and electrical gain analysis using a finite-element method. Mechanical displacement and electric potential of the transformer at the calculated resonance frequency are used to calculate the loss distribution of the transformer. Temperature distribution using discretized heat transfer equation is calculated from the obtained losses of the transformer. Properties of the piezoelectric material, dependent on the temperature field, are measured to recalculate the losses, temperature distribution, and new resonance characteristics of the transformer. Iterative method is adopted to recalculate the losses and resonance frequency due to the changes of the material constants from temperature increase. Computed temperature distributions and new resonance characteristics of the transformer at steady-state temperature are verified by comparison with experimental results.

  18. Apparatus and methods for continuous beam fourier transform mass spectrometry

    DOEpatents

    McLuckey, Scott A.; Goeringer, Douglas E.

    2002-01-01

    A continuous beam Fourier transform mass spectrometer in which a sample of ions to be analyzed is trapped in a trapping field, and the ions in the range of the mass-to-charge ratios to be analyzed are excited at their characteristic frequencies of motion by a continuous excitation signal. The excited ions in resonant motions generate real or image currents continuously which can be detected and processed to provide a mass spectrum.

  19. Modeling of a method of parallel hierarchical transformation for fast recognition of dynamic images

    NASA Astrophysics Data System (ADS)

    Timchenko, Leonid I.; Kokryatskaya, Nataliya I.; Shpakovych, Viktoriya V.

    2013-12-01

    Principles necessary to develop a method and computational facilities for the parallel hierarchical transformation based on high-performance GPUs are discussed in the paper. Mathematic models of the parallel hierarchical (PH) network training for the transformation and a PH network training method for recognition of dynamic images are developed.

  20. A Unified Method of Finding Laplace Transforms, Fourier Transforms, and Fourier Series. [and] An Inversion Method for Laplace Transforms, Fourier Transforms, and Fourier Series. Integral Transforms and Series Expansions. Modules and Monographs in Undergraduate Mathematics and Its Applications Project. UMAP Units 324 and 325.

    ERIC Educational Resources Information Center

    Grimm, C. A.

    This document contains two units that examine integral transforms and series expansions. In the first module, the user is expected to learn how to use the unified method presented to obtain Laplace transforms, Fourier transforms, complex Fourier series, real Fourier series, and half-range sine series for given piecewise continuous functions. In…

  1. On the Inclusion of Difference Equation Problems and Z Transform Methods in Sophomore Differential Equation Classes

    ERIC Educational Resources Information Center

    Savoye, Philippe

    2009-01-01

    In recent years, I started covering difference equations and z transform methods in my introductory differential equations course. This allowed my students to extend the "classical" methods for (ordinary differential equation) ODE's to discrete time problems arising in many applications.

  2. Maize transformation technology development for commercial event generation

    PubMed Central

    Que, Qiudeng; Elumalai, Sivamani; Li, Xianggan; Zhong, Heng; Nalapalli, Samson; Schweiner, Michael; Fei, Xiaoyin; Nuccio, Michael; Kelliher, Timothy; Gu, Weining; Chen, Zhongying; Chilton, Mary-Dell M.

    2014-01-01

    Maize is an important food and feed crop in many countries. It is also one of the most important target crops for the application of biotechnology. Currently, there are more biotech traits available on the market in maize than in any other crop. Generation of transgenic events is a crucial step in the development of biotech traits. For commercial applications, a high throughput transformation system producing a large number of high quality events in an elite genetic background is highly desirable. There has been tremendous progress in Agrobacterium-mediated maize transformation since the publication of the Ishida et al. (1996) paper and the technology has been widely adopted for transgenic event production by many labs around the world. We will review general efforts in establishing efficient maize transformation technologies useful for transgenic event production in trait research and development. The review will also discuss transformation systems used for generating commercial maize trait events currently on the market. As the number of traits is increasing steadily and two or more modes of action are used to control key pests, new tools are needed to efficiently transform vectors containing multiple trait genes. We will review general guidelines for assembling binary vectors for commercial transformation. Approaches to increase transformation efficiency and gene expression of large gene stack vectors will be discussed. Finally, recent studies of targeted genome modification and transgene insertion using different site-directed nuclease technologies will be reviewed. PMID:25140170

  3. Hilbert-Huang transformation-based time-frequency analysis methods in biomedical signal applications.

    PubMed

    Lin, Chin-Feng; Zhu, Jin-De

    2012-03-01

    Hilbert-Huang transformation, wavelet transformation, and Fourier transformation are the principal time-frequency analysis methods. These transformations can be used to discuss the frequency characteristics of linear and stationary signals, the time-frequency features of linear and non-stationary signals, the time-frequency features of non-linear and non-stationary signals, respectively. The Hilbert-Huang transformation is a combination of empirical mode decomposition and Hilbert spectral analysis. The empirical mode decomposition uses the characteristics of signals to adaptively decompose them to several intrinsic mode functions. Hilbert transforms are then used to transform the intrinsic mode functions into instantaneous frequencies, to obtain the signal's time-frequency-energy distributions and features. Hilbert-Huang transformation-based time-frequency analysis can be applied to natural physical signals such as earthquake waves, winds, ocean acoustic signals, mechanical diagnosis signals, and biomedical signals. In previous studies, we examined Hilbert-Huang transformation-based time-frequency analysis of the electroencephalogram FPI signals of clinical alcoholics, and 'sharp I' wave-based Hilbert-Huang transformation time-frequency features. In this paper, we discuss the application of Hilbert-Huang transformation-based time-frequency analysis to biomedical signals, such as electroencephalogram, electrocardiogram signals, electrogastrogram recordings, and speech signals.

  4. Mechanic-electrical transformations in the Kelvin method

    NASA Astrophysics Data System (ADS)

    Zharkikh, Yu. S.; Lysochenko, S. V.

    2017-04-01

    To explain the initiation mechanism of alternating current in an electric circuit containing the dynamic capacitor a model of mechanic- electrical transformation is suggested to use. In such a model, electric charges disposed between the capacitor plates serve as a cause of measured signal in contrast to the contact potential difference, which is considered as the main base in the Kelvin's model. If one of the plates moves periodically, then the conditions of the charges screening are changed and thereby the capacitor recharging current is arise. The measuring is based on compensation of the recharging current by current, which generated by a source of electromotive force (EMF). The compensation voltage depends on both the distribution of ions or dipoles over the studied surface and the charges creating the surface potential barrier. This voltage is independent on the bulk electro-physical characteristics of a solid.

  5. Issues and progress in transforming a middle-division classical mechanics/math methods course

    NASA Astrophysics Data System (ADS)

    Pollock, Steven J.; Pepper, Rachel E.; Marino, Alysia D.

    2012-02-01

    The physics department at the University of Colorado, Boulder has recently begun the transformation of its Classical Mechanics/Math Methods course, a middle-division course taken primarily by sophomore physics majors. We discuss the process of course transformation, including holding faculty meetings to create consensus learning goals and a conceptual diagnostic, and adopting, adapting and creating course materials and structures. We also report preliminary observations of student learning gains, student attitudes towards the transformation, and common student difficulties with the course material. We also discuss ongoing plans for the course transformation.

  6. Remote sensing image compression method based on lift scheme wavelet transform

    NASA Astrophysics Data System (ADS)

    Tao, Hongjiu; Tang, Xinjian; Liu, Jian; Tian, Jinwen

    2003-06-01

    Based on lifting scheme and the construction theorem of the integer Haar wavelet and biorthogonal wavelet, we propose a new integer wavelet transform construct method on the basis of lift scheme after introduciton of constructing specific-demand biorthogonal wavelet transform using Harr wavelet and Lazy wavelet. In this paper, we represent the method and algorithm of the lifting scheme, and we also give mathematical formulation on this method and experimental results as well.

  7. Design of piezoelectric transformer for DC/DC converter with stochastic optimization method

    NASA Astrophysics Data System (ADS)

    Vasic, Dejan; Vido, Lionel

    2016-04-01

    Piezoelectric transformers were adopted in recent year due to their many inherent advantages such as safety, no EMI problem, low housing profile, and high power density, etc. The characteristics of the piezoelectric transformers are well known when the load impedance is a pure resistor. However, when piezoelectric transformers are used in AC/DC or DC/DC converters, there are non-linear electronic circuits connected before and after the transformer. Consequently, the output load is variable and due to the output capacitance of the transformer the optimal working point change. This paper starts from modeling a piezoelectric transformer connected to a full wave rectifier in order to discuss the design constraints and configuration of the transformer. The optimization method adopted here use the MOPSO algorithm (Multiple Objective Particle Swarm Optimization). We start with the formulation of the objective function and constraints; then the results give different sizes of the transformer and the characteristics. In other word, this method is looking for a best size of the transformer for optimal efficiency condition that is suitable for variable load. Furthermore, the size and the efficiency are found to be a trade-off. This paper proposes the completed design procedure to find the minimum size of PT in need. The completed design procedure is discussed by a given specification. The PT derived from the proposed design procedure can guarantee both good efficiency and enough range for load variation.

  8. Commentary: Using Mixed Methods to Transform Special Education Research

    ERIC Educational Resources Information Center

    Trainor, Audrey A.

    2011-01-01

    Klingner and Boardman (this issue) offer a cogent and compelling argument for opening the door for the acceptance and use of mixed methods in special education research. Self-identifying as pragmatists, they embody this paradigmatic view by focusing on the utility, efficacy, and accuracy of mixed methods, an argument that should appeal to the…

  9. Efficient genetic transformation of red raspberry, Rubus ideaus L.

    PubMed

    Mathews, H; Wagoner, W; Cohen, C; Kellogg, J; Bestwick, R

    1995-05-01

    We have developed an efficient transformation system for red raspberry (Rubus ideaus L.) using Agrobacterium mediated gene transfer. Using this system we have successfully introduced a gene that encodes an enzyme, S-adenosylmethionine hydrolase (SAMase), in raspberry cultivars Meeker (MK), Chilliwack (CH) and Canby (CY). Leaf and petiole expiants were inoculated with disarmed Agrobacterium tumefaciens strain EHA 105 carrying either of two binary vectors, pAG1452 or pAG1552, encoding gene sequences for SAMase under the control of the wound and fruit specific tomato E4 promoter. Primary shoot regenerants on selection medium were chimeral containing both transformed and non-transformed cells. Non-chimeral transgenic clones were developed by iterative culture of petiole, node and leaf explants, on selection medium, from successive generations of shoots derived from the primary regenerants. Percent recovery of transformants was higher with the selection marker gene hygromycin phosphotransferase (hpt), than with neomycin phosphotransferase (nptII). Transformation frequencies of 49.6%, 0.9% and 8.1% were obtained in cultivars Meeker, Chilliwack and Canby respectively from petiole expiants using hygromycin selection. Genomic integration of transgenes was confirmed by Southern hybridization. Transgenic plants from a total of 218 independent transformation events (161 MK, 4 CH, 53 CY) have been successfully established in soil.

  10. Transforming student's discourse as a method of teaching science inquiry

    NASA Astrophysics Data System (ADS)

    Livingston, David

    2005-07-01

    A qualitative case study on the instructional practice of one secondary science teacher addresses the persistent reluctance of many science teachers to integrate the cultural resources and social practices of professional science communities into the science content they teach. The literature has shown that teachers' hesitation to implement a social and locally situated learning strategy curtails students' ability to draw upon the language of science necessary to co-construct and shape authentic science inquiry and in particular appropriate argument schemes. The study hypothesized that a teacher's dialogic facilitation of a particular social context and instructional practices enhances a students' ability to express verbally the claims and warrants that rise from evidence taken from their inquiries of natural phenomena. The study also tracks students' use of the Key Words and Ideas of this science curriculum for the purpose of assessing the degree of students' assimilation of these terms into their speech and written expressions of inquiry. The theoretical framework is Vygotskian (1978) and the analysis of the qualitative data is founded on Toulmin (1958), Walton (1996), Jimenez-Alexandre et al. (2000) and Shavelson (1996). The dialogic structure of this teacher's facilitation of student's science knowledge is shown to utilize students' presumptive statements to hone their construction of inductive or deductive arguments. This instructional practice may represent teacher-student activity within the zone of proximal development and supports Vygotsky's notion that a knowledgeable other is instrumental in transforming student's spontaneous talk into scientific speech. The tracking of the curriculum's Key Words and Ideas into students' speech and writing indicated that this teachers' ability to facilitate students' presumptuous reasoning into logic statements did not necessarily guarantee that they could post strong written expressions of this verbal know-how in

  11. Missing texture reconstruction method based on error reduction algorithm using Fourier transform magnitude estimation scheme.

    PubMed

    Ogawa, Takahiro; Haseyama, Miki

    2013-03-01

    A missing texture reconstruction method based on an error reduction (ER) algorithm, including a novel estimation scheme of Fourier transform magnitudes is presented in this brief. In our method, Fourier transform magnitude is estimated for a target patch including missing areas, and the missing intensities are estimated by retrieving its phase based on the ER algorithm. Specifically, by monitoring errors converged in the ER algorithm, known patches whose Fourier transform magnitudes are similar to that of the target patch are selected from the target image. In the second approach, the Fourier transform magnitude of the target patch is estimated from those of the selected known patches and their corresponding errors. Consequently, by using the ER algorithm, we can estimate both the Fourier transform magnitudes and phases to reconstruct the missing areas.

  12. Modified variational iteration method for partial differential equations using Ma's transformation

    NASA Astrophysics Data System (ADS)

    Mohyud-Din, Syed Tauseef

    2010-03-01

    In this paper, we apply the modified variational iteration method (MVIM) for solving partial differential equations using Ma's transformation. The proposed modification is made by introducing He's polynomials in the correction functional of the variational iteration method (VIM). Moreover, we use a very efficient and reliable transformation which is mainly due to Ma and converts the given partial differential equations (PDES) into the corresponding ordinary differential equations (ODES). The proposed MVIM is applied on the re-formulated ODES, which in turn gives the solution in terms of the transformed variables and the application of the inverse transformation yields the required series solution. Several examples are given to re-confirm the efficiency and accuracy of the suggested algorithm. It is observed that Ma's transformation makes the solution procedure very convenient and simple.

  13. Agrobacterium tumefaciens-mediated transformation of poinsettia, Euphorbia pulcherrima, with virus-derived hairpin RNA constructs confers resistance to Poinsettia mosaic virus.

    PubMed

    Clarke, Jihong Liu; Spetz, Carl; Haugslien, Sissel; Xing, Shaochen; Dees, Merete W; Moe, Roar; Blystad, Dag-Ragnar

    2008-06-01

    Agrobacterium-mediated transformation for poinsettia (Euphorbia pulcherrima Willd. Ex Klotzsch) is reported here for the first time. Internode stem explants of poinsettia cv. Millenium were transformed by Agrobacterium tumefaciens, strain LBA 4404, harbouring virus-derived hairpin (hp) RNA gene constructs to induce RNA silencing-mediated resistance to Poinsettia mosaic virus (PnMV). Prior to transformation, an efficient somatic embryogenesis system was developed for poinsettia cv. Millenium in which about 75% of the explants produced somatic embryos. In 5 experiments utilizing 868 explants, 18 independent transgenic lines were generated. An average transformation frequency of 2.1% (range 1.2-3.5%) was revealed. Stable integration of transgenes into the poinsettia nuclear genome was confirmed by PCR and Southern blot analysis. Both single- and multiple-copy transgene integration into the poinsettia genome were found among transformants. Transgenic poinsettia plants showing resistance to mechanical inoculation of PnMV were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Northern blot analysis of low molecular weight RNA revealed that transgene-derived small interfering (si) RNA molecules were detected among the poinsettia transformants prior to inoculation. The Agrobacterium-mediated transformation methodology developed in the current study should facilitate improvement of this ornamental plant with enhanced disease resistance, quality improvement and desirable colour alteration. Because poinsettia is a non-food, non-feed plant and is not propagated through sexual reproduction, this is likely to be more acceptable even in areas where genetically modified crops are currently not cultivated.

  14. Transformational Teaching: Theoretical Underpinnings, Basic Principles, and Core Methods

    ERIC Educational Resources Information Center

    Slavich, George M.; Zimbardo, Philip G.

    2012-01-01

    Approaches to classroom instruction have evolved considerably over the past 50 years. This progress has been spurred by the development of several learning principles and methods of instruction, including active learning, student-centered learning, collaborative learning, experiential learning, and problem-based learning. In the present paper, we…

  15. The first transformation method for the thermo-acidophilic archaeon Thermoplasma acidophilum.

    PubMed

    Baka, Erzsébet; Varga, Sándor; Hobel, Cédric; Knispel, Roland Wilhelm; Fekete, Csaba; Ivanics, Milán; Kriszt, Balázs; Nagy, István; Kukolya, József

    2013-11-01

    A transformation method yielding up to 10(4) transformants per μg circular DNA was developed for Thermoplasma acidophilum. The method is based on a natural DNA uptake process in which T. acidophilum cells keep their integrity and turn competent at pH 3.5 and 58°C. Shuttle vector maintenance could not be detected, since the used Nov(R) gyraseB gene integrated into its chromosomal counterpart by homologous recombination.

  16. A Rapid Coordinate Transformation Method Applied in Industrial Robot Calibration Based on Characteristic Line Coincidence.

    PubMed

    Liu, Bailing; Zhang, Fumin; Qu, Xinghua; Shi, Xiaojia

    2016-02-18

    Coordinate transformation plays an indispensable role in industrial measurements, including photogrammetry, geodesy, laser 3-D measurement and robotics. The widely applied methods of coordinate transformation are generally based on solving the equations of point clouds. Despite the high accuracy, this might result in no solution due to the use of ill conditioned matrices. In this paper, a novel coordinate transformation method is proposed, not based on the equation solution but based on the geometric transformation. We construct characteristic lines to represent the coordinate systems. According to the space geometry relation, the characteristic line scan is made to coincide by a series of rotations and translations. The transformation matrix can be obtained using matrix transformation theory. Experiments are designed to compare the proposed method with other methods. The results show that the proposed method has the same high accuracy, but the operation is more convenient and flexible. A multi-sensor combined measurement system is also presented to improve the position accuracy of a robot with the calibration of the robot kinematic parameters. Experimental verification shows that the position accuracy of robot manipulator is improved by 45.8% with the proposed method and robot calibration.

  17. FFT-enhanced IHS transform method for fusing high-resolution satellite images

    USGS Publications Warehouse

    Ling, Y.; Ehlers, M.; Usery, E.L.; Madden, M.

    2007-01-01

    Existing image fusion techniques such as the intensity-hue-saturation (IHS) transform and principal components analysis (PCA) methods may not be optimal for fusing the new generation commercial high-resolution satellite images such as Ikonos and QuickBird. One problem is color distortion in the fused image, which causes visual changes as well as spectral differences between the original and fused images. In this paper, a fast Fourier transform (FFT)-enhanced IHS method is developed for fusing new generation high-resolution satellite images. This method combines a standard IHS transform with FFT filtering of both the panchromatic image and the intensity component of the original multispectral image. Ikonos and QuickBird data are used to assess the FFT-enhanced IHS transform method. Experimental results indicate that the FFT-enhanced IHS transform method may improve upon the standard IHS transform and the PCA methods in preserving spectral and spatial information. ?? 2006 International Society for Photogrammetry and Remote Sensing, Inc. (ISPRS).

  18. A Rapid Coordinate Transformation Method Applied in Industrial Robot Calibration Based on Characteristic Line Coincidence

    PubMed Central

    Liu, Bailing; Zhang, Fumin; Qu, Xinghua; Shi, Xiaojia

    2016-01-01

    Coordinate transformation plays an indispensable role in industrial measurements, including photogrammetry, geodesy, laser 3-D measurement and robotics. The widely applied methods of coordinate transformation are generally based on solving the equations of point clouds. Despite the high accuracy, this might result in no solution due to the use of ill conditioned matrices. In this paper, a novel coordinate transformation method is proposed, not based on the equation solution but based on the geometric transformation. We construct characteristic lines to represent the coordinate systems. According to the space geometry relation, the characteristic line scan is made to coincide by a series of rotations and translations. The transformation matrix can be obtained using matrix transformation theory. Experiments are designed to compare the proposed method with other methods. The results show that the proposed method has the same high accuracy, but the operation is more convenient and flexible. A multi-sensor combined measurement system is also presented to improve the position accuracy of a robot with the calibration of the robot kinematic parameters. Experimental verification shows that the position accuracy of robot manipulator is improved by 45.8% with the proposed method and robot calibration. PMID:26901203

  19. Efficient regeneration and improved sonication-assisted Agrobacterium transformation (SAAT) method for Catharanthus roseus.

    PubMed

    Alam, Pravej; Khan, Zainul Abdeen; Abdin, Malik Zainul; Khan, Jawaid A; Ahmad, Parvaiz; Elkholy, Shereen F; Sharaf-Eldin, Mahmoud A

    2017-05-01

    Catharanthus roseus is an important medicinal plant known for its pharmacological qualities such as antimicrobial, anticancerous, antifeedant, antisterility, antidiabetic activities. More than 130 bioactive compounds like vinblastine, vindoline and vincristine have been synthesized in this plant. Extensive studies have been carried out for optimization regeneration and transformation protocols. Most of the protocol described are laborious and time-consuming. Due to sophisticated protocol of regeneration and genetic transformation, the production of these bioactive molecules is less and not feasible to be commercialized worldwide. Here we have optimized the efficient protocol for regeneration and transformation to minimize the time scale and enhance the transformation frequency through Agrobacterium and sonication-assisted transformation (SAAT) method. In this study, hypocotyl explants responded best for maximal production of transformed shoots. The callus percentage were recorded 52% with 1.0 mg L(-1) (BAP) and 0.5 mg L(-1) (NAA) while 80% shoot percentage obtained with 4.0 mg L(-1) (BAP) and 0.05 mg L(-1) (NAA). The microscopic studies revealed that the expression of GFP was clearly localized in leaf tissue of the C. roseus after transformation of pRepGFP0029 construct. Consequently, transformation efficiency was revealed on the basis of GFP localization. The transformation efficiency of SAAT method was 6.0% comparable to 3.5% as conventional method. Further, PCR analysis confirmed the integration of the nptII gene in the transformed plantlets of C. roseus.

  20. Methods for transforming and expression screening of filamentous fungal cells with a DNA library

    DOEpatents

    Teter, Sarah; Lamsa, Michael; Cherry, Joel; Ward, Connie

    2015-06-02

    The present invention relates to methods for expression screening of filamentous fungal transformants, comprising: (a) isolating single colony transformants of a DNA library introduced into E. coli; (b) preparing DNA from each of the single colony E. coli transformants; (c) introducing a sample of each of the DNA preparations of step (b) into separate suspensions of protoplasts of a filamentous fungus to obtain transformants thereof, wherein each transformant contains one or more copies of an individual polynucleotide from the DNA library; (d) growing the individual filamentous fungal transformants of step (c) on selective growth medium, thereby permitting growth of the filamentous fungal transformants, while suppressing growth of untransformed filamentous fungi; and (e) measuring activity or a property of each polypeptide encoded by the individual polynucleotides. The present invention also relates to isolated polynucleotides encoding polypeptides of interest obtained by such methods, to nucleic acid constructs, expression vectors, and recombinant host cells comprising the isolated polynucleotides, and to methods of producing the polypeptides encoded by the isolated polynucleotides.

  1. A method to analyze molecular tagging velocimetry data using the Hough transform.

    PubMed

    Sanchez-Gonzalez, R; McManamen, B; Bowersox, R D W; North, S W

    2015-10-01

    The development of a method to analyze molecular tagging velocimetry data based on the Hough transform is presented. This method, based on line fitting, parameterizes the grid lines "written" into a flowfield. Initial proof-of-principle illustration of this method was performed to obtain two-component velocity measurements in the wake of a cylinder in a Mach 4.6 flow, using a data set derived from computational fluid dynamics simulations. The Hough transform is attractive for molecular tagging velocimetry applications since it is capable of discriminating spurious features that can have a biasing effect in the fitting process. Assessment of the precision and accuracy of the method were also performed to show the dependence on analysis window size and signal-to-noise levels. The accuracy of this Hough transform-based method to quantify intersection displacements was determined to be comparable to cross-correlation methods. The employed line parameterization avoids the assumption of linearity in the vicinity of each intersection, which is important in the limit of drastic grid deformations resulting from large velocity gradients common in high-speed flow applications. This Hough transform method has the potential to enable the direct and spatially accurate measurement of local vorticity, which is important in applications involving turbulent flowfields. Finally, two-component velocity determinations using the Hough transform from experimentally obtained images are presented, demonstrating the feasibility of the proposed analysis method.

  2. Blind Forensics of Successive Geometric Transformations in Digital Images using Spectral Method: Theory and Applications.

    PubMed

    Ni, Jiangqun; Chen, Chenglong; Shen, Zhaoyi; Shi, Yun-Qing

    2017-03-15

    Geometric transformations, such as resizing and rotation, are almost always needed when two or more images are spliced together to create convincing image forgeries. In recent years, researchers have developed many digital forensic techniques to identify these operations. Most previous works in this area focus on the analysis of images that have undergone single geometric transformations, e.g., resizing or rotation. In several recent works, researchers have addressed yet another practical and realistic situation: successive geometric transformations, e.g., repeated resizing, resizing-rotation, rotation-resizing, and repeated rotation. We will also concentrate on this topic in this paper. Specifically, we present an in-depth analysis in the frequency domain of the second-order statistics of the geometrically transformed images. We give an exact formulation of how the parameters of the first and second geometric transformations influence the appearance of periodic artifacts. The expected positions of characteristic resampling peaks are analytically derived. The theory developed here helps to address the gap left by previous works on this topic and is useful for image security and authentication, in particular, the forensics of geometric transformations in digital images. As an application of the developed theory, we present an effective method that allows one to distinguish between the aforementioned four different processing chains. The proposed method can further estimate all the geometric transformation parameters. This may provide useful clues for image forgery detection.

  3. Modal wavefront estimation from its slopes by numerical orthogonal transformation method over general shaped aperture.

    PubMed

    Ye, Jingfei; Wang, Wei; Gao, Zhishan; Liu, Zhiying; Wang, Shuai; Benítez, Pablo; Miñano, Juan C; Yuan, Qun

    2015-10-05

    Wavefront estimation from the slope-based sensing metrologies zis important in modern optical testing. A numerical orthogonal transformation method is proposed for deriving the numerical orthogonal gradient polynomials as numerical orthogonal basis functions for directly fitting the measured slope data and then converting to the wavefront in a straightforward way in the modal approach. The presented method can be employed in the wavefront estimation from its slopes over the general shaped aperture. Moreover, the numerical orthogonal transformation method could be applied to the wavefront estimation from its slope measurements over the dynamic varying aperture. The performance of the numerical orthogonal transformation method is discussed, demonstrated and verified by the examples. They indicate that the presented method is valid, accurate and easily implemented for wavefront estimation from its slopes.

  4. Practical Symplectic Methods with Time Transformation for the Few-Body Problem

    NASA Astrophysics Data System (ADS)

    Mikkola, Seppo

    1997-02-01

    The use of the extended phase space and time transformations for constructing efficient symplectic algorithms for the investigation of long term behavior of hierarchical few-body systems is discussed. Numerical experiments suggest that the time-transformed generalized leap-frog, combined with symplectic correctors, is one of the most efficient methods for such studies. Applications extend from perturbed two-body motion to hierarchical many-body systems with large eccentricities.

  5. A Transformer Coupling Method for Imaging Defects in Concentrically Arranged Steel Tubes

    SciTech Connect

    Miller, G.; Hussin, H.; Fernandes, B.; Zaid, M.; Gaydecki, P.; El-Madaani, F.

    2006-03-06

    This paper describes the development of a system that uses a transformer coupling method for imaging defects in the outer tube of a concentric steel tube pair. Transformer coupling is employed where a receiver is mounted opposite to the transmitter. Magnetic field coupling is used, in which the field propagates through the inner steel tube towards the outer steel tube. Defects in the outer section distort the field and the change is detected by the receiver.

  6. Design of high-performance fixed-point transforms using the common factor method

    NASA Astrophysics Data System (ADS)

    Hinds, Arianne T.

    2010-08-01

    Fixed-point implementations of transforms such as the Discrete Cosine Transform (DCT) remain as fundamental building blocks of state-of-the-art video coding technologies. Recently, the 16x16 DCT has received focus as a transform suitable for the high efficiency video coding project currently underway in the Joint Collaboration Team - Video Coding. By its definition, the 16x16 DCT is inherently more complex than transforms of traditional sizes such as 4x4 or 8x8 DCTs. However, scaled architectures such as the one employed in the design of the 8x8 DCTs specified in ISO/IEC 23002-2 can also be utilized to mitigate the complexity of fixed-point approximations of higher-order transforms such as the 16x16 DCT. This paper demonstrates the application of the Common Factor method to design two scaled implementations of the 16x16 DCT. One implementation can be characterized by its exceptionally low complexity, while the other can be characterized by its relatively high precision. We review the Common Factor method as a method to arrive at fixed-point implementations that are optimized in terms of complexity and precision for such high performance transforms.

  7. A Simple and Non-Invasive Method for Nuclear Transformation of Intact-walled Chlamydomonas reinhardtii

    PubMed Central

    Kim, Sora; Lee, Young-Chul; Cho, Dae-Hyun; Lee, Hyun Uk; Huh, Yun Suk; Kim, Geun-Joong; Kim, Hee-Sik

    2014-01-01

    Genetic engineering in microalgae is gaining attraction but nuclear transformation methods available so far are either inefficient or require special equipment. In this study, we employ positively charged nanoparticles, 3-aminopropyl-functionalized magnesium phyllosilicate (aminoclay, approximate unit cell composition of [H2N(CH2)3]8Si8Mg6O12(OH)4), for nuclear transformation into eukaryotic microalgae. TEM and EDX analysis of the process of transformation reveals that aminoclay coats negatively-charged DNA biomolecules and forms a self-assembled hybrid nanostructure. Subsequently, when this nanostructure is mixed with microalgal cells and plated onto selective agar plates with high friction force, cell wall is disrupted facilitating delivery of plasmid DNA into the cell and ultimately to the nucleus. This method is not only simple, inexpensive, and non-toxic to cells but also provides efficient transformation (5.03×102 transformants/µg DNA), second only to electroporation which needs advanced instrumentation. We present optimized parameters for efficient transformation including pre-treatment, friction force, concentration of foreign DNA/aminoclay, and plasticity of agar plates. It is also confirmed the successful integration and stable expression of foreign gene in Chlamydomonas reinhardtii through molecular methods. PMID:24988123

  8. Basic Investigation on Medical Ultrasonic Echo Image Compression by JPEG2000 - Availability of Wavelet Transform and ROI Method

    DTIC Science & Technology

    2007-11-02

    be approved in the near future. The main features of JPEG2000 are use of wavelet transform and ROI (Region of Interest) method. It is expected that... wavelet transform is more effective than Fourier transform for ultrasonic echo signal/image processing. Furthermore, ROI method seems to be appropriate...compression method of medical images. The purpose of this paper is to investigate the effectiveness of wavelet transform compared with DCT (JPEG) and

  9. [Transformation of Didymella bryoniae mediated by Agrobacterium tumefaciens].

    PubMed

    Ren, Haiying; Fang, Li; Li, Gang; Ru, Shuijiang; Wang, Hanrong

    2010-06-01

    Gummy stem blight, a plant disease caused by Didymella bryoniae, is one of the major diseases in melon. The disease can seriously reduce melon yield and quality. However, little information is available on the genetics and functional genomics of the fungal pathogen. In this study, we developed an Agrobacterium-mediated transformation system for D. bryoniae by using a universal pathogenic isolate DB11 and the Agrobacterium tumefaciens strain C58C1 carrying plasmid pBIG2RHPH2 harboring the hygromycin B phosphotransferase gene (hph). Total 45 transformants could be obtained per 1 x 10(5) spores when 1 x 10(6) spores per milliliter of D. bryoniae spore suspension were cocultivated with Agrobacterium cells at OD600 = 0.15 for 48 h in the presence of induction medium (pH 5.2) containing acetosyringone at 200 microg/mL and selection medium contained 100 microg/mL of hygromycin B and 200 microg/mL of cefotaxime sodium, ampicillin and tetracycline, respectively. The transformants were stable when grown on PDA medium without hygromycin B for five times and were verified by PCR amplification with the hph primers and by Southern blot analysis with the hph probe. The transformation system will be useful for further studies of functional genes in D. bryoniae.

  10. Test of the method for calculation of derating of workshop transformers on engineering plants

    NASA Astrophysics Data System (ADS)

    Shklyarskiy, A. Y.; Bardanov, A. I.

    2017-02-01

    Today’s innovations in engineering production are aimed at the improvement of equipment efficiency in order to reduce losses and to save environment. It is conventional that the adjustable-speed drive improves electrical and mechanical transients of workshop units, but power electronics’ devices are current harmonics sources. Presence of high harmonic currents in the workshop’s network is a substantial reason for transformer derating. The article discloses the method approbation, the method for calculating additional losses in an idle transformer for non-sine feeding voltage. The approbation was carried out via comparing the calculated data with the experimental data. The article describes the studies method, its strong and weak sides, and it also presents the actual calculation of coefficients for a math model, when calculating losses in a transformer, based on existing experimental data. The conclusion comprises the observation regarding possibility to use the method described to solve practical tasks.

  11. Chemometric Analysis of Multicomponent Biodegradable Plastics by Fourier Transform Infrared Spectrometry: The R-Matrix Method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A new chemometric method based on absorbance ratios from Fourier transform infrared spectra was devised to analyze multicomponent biodegradable plastics. The method uses the BeerLambert law to directly compute individual component concentrations and weight losses before and after biodegradation of c...

  12. A new rapid method for quantification of PCBs in transformer oil.

    PubMed

    Takada, M; Toda, H; Uchida, R

    2001-01-01

    In order to improve the efficiency and cost-effectiveness of the PCB analysis, the DMSO partition and SPE extraction were applied to clean up the PCB-contaminated transformer oils and PCB level was determined by means of the quadrupole GC/MS. The analysis data obtained from this method were compared statistically to that from the standard method.

  13. Knowledge Transmission versus Social Transformation: A Critical Analysis of Purpose in Elementary Social Studies Methods Textbooks

    ERIC Educational Resources Information Center

    Butler, Brandon M.; Suh, Yonghee; Scott, Wendy

    2015-01-01

    In this article, the authors investigate the extent to which 9 elementary social studies methods textbooks present the purpose of teaching and learning social studies. Using Stanley's three perspectives of teaching social studies for knowledge transmission, method of intelligence, and social transformation; we analyze how these texts prepare…

  14. [Comparison of correction methods for nonlinear optic path difference of reflecting rotating Fourier transform spectrometer].

    PubMed

    Jing, Juan-Juan; Zhou, Jin-Song; Xiangli, Bin; Lü, Qun-Bo; Wei, Ru-Yi

    2010-06-01

    The principle of reflecting rotating Fourier transform spectrometer was introduced in the present paper. The nonlinear problem of optical path difference (OPD) of rotating Fourier transform spectrometer universally exists, produced by the rotation of rotating mirror. The nonlinear OPD will lead to fictitious recovery spectrum, so it is necessary to compensate the nonlinear OPD. Three methods of correction for the nonlinear OPD were described and compared in this paper, namely NUFFT method, OPD replace method and interferograms fitting method. The result indicates that NUFFT was the best method for the compensation of nonlinear OPD, OPD replace method was better, its precision was almost the same as NUFFT method, and their relative error are superior to 0.13%, but the computation efficiency of OPD replace method is slower than NUFFT method, while the precision and computation efficiency of interferograms fitting method are not so satisfied, because the interferograms are rapid fluctuant especially around the zero optical path difference, so it is unsuitable for polynomial fitting, and because this method needs piecewise fitting, its computation efficiency is the slowest, thus the NUFFT method is the most suited method for the nonlinear OPD compensation of reflecting rotating Fourier transform spectrometer.

  15. Genetic transformation of cotton with a harpin-encoding gene hpaXoo confers an enhanced defense response against Verticillium dahliae Kleb.

    PubMed

    Miao, Weiguo; Wang, Jingsheng

    2013-01-01

    The soil-borne fungal pathogen Verticillium dahliae Kleb causes Verticillium wilt in a wide range of crops including cotton (Gossypium hirsutum). To date, most upland cotton varieties are susceptible to V. dahliae and the breeding for cotton varieties with the resistance to Verticillium wilt has not been successful. Hpa1Xoo is a harpin protein from Xanthomonas oryzae pv. oryzae which induces the hypersensitive cell death in plants. When hpa1Xoo was transformed into the susceptible cotton line Z35 through Agrobacterium-mediated transformation, the transgenic cotton line (T-34) with an improved resistance to Verticillium dahliae was obtained. Here, we describe the related research approach, such as Western blot, Southern blot, immuno-gold labeling, evaluation of resistance to Verticillium dahliae, and how to detect the micro-hypersensitive response and oxidative burst elicited by harpin(Xoo) in plant tissue.

  16. The method of parallel-hierarchical transformation for rapid recognition of dynamic images using GPGPU technology

    NASA Astrophysics Data System (ADS)

    Timchenko, Leonid; Yarovyi, Andrii; Kokriatskaya, Nataliya; Nakonechna, Svitlana; Abramenko, Ludmila; Ławicki, Tomasz; Popiel, Piotr; Yesmakhanova, Laura

    2016-09-01

    The paper presents a method of parallel-hierarchical transformations for rapid recognition of dynamic images using GPU technology. Direct parallel-hierarchical transformations based on cluster CPU-and GPU-oriented hardware platform. Mathematic models of training of the parallel hierarchical (PH) network for the transformation are developed, as well as a training method of the PH network for recognition of dynamic images. This research is most topical for problems on organizing high-performance computations of super large arrays of information designed to implement multi-stage sensing and processing as well as compaction and recognition of data in the informational structures and computer devices. This method has such advantages as high performance through the use of recent advances in parallelization, possibility to work with images of ultra dimension, ease of scaling in case of changing the number of nodes in the cluster, auto scan of local network to detect compute nodes.

  17. Quantitative interferometric microscopy with two dimensional Hilbert transform based phase retrieval method

    NASA Astrophysics Data System (ADS)

    Wang, Shouyu; Yan, Keding; Xue, Liang

    2017-01-01

    In order to obtain high contrast images and detailed descriptions of label free samples, quantitative interferometric microscopy combining with phase retrieval is designed to obtain sample phase distributions from fringes. As accuracy and efficiency of recovered phases are affected by phase retrieval methods, thus approaches owning higher precision and faster processing speed are still in demand. Here, two dimensional Hilbert transform based phase retrieval method is adopted in cellular phase imaging, it not only reserves more sample specifics compared to classical fast Fourier transform based method, but also overcomes disadvantages of traditional algorithm according to Hilbert transform which is a one dimensional processing causing phase ambiguities. Both simulations and experiments are provided, proving the proposed phase retrieval approach can acquire quantitative sample phases with high accuracy and fast speed.

  18. Performance comparison of ISAR imaging method based on time frequency transforms

    NASA Astrophysics Data System (ADS)

    Xie, Chunjian; Guo, Chenjiang; Xu, Jiadong

    2013-03-01

    Inverse synthetic aperture radar (ISAR) can image the moving target, especially the target in the air, so it is important in the air defence and missile defence system. Time-frequency Transform was applied to ISAR imaging process widely. Several time frequency transforms were introduced. Noise jamming methods were analysed, and when these noise jamming were added to the echo of the ISAR receiver, the image can become blur even can't to be identify. But the effect is different to the different time frequency analysis. The results of simulation experiment show the Performance Comparison of the method.

  19. Use of ultrasonic array method for positioning multiple partial discharge sources in transformer oil.

    PubMed

    Xie, Qing; Tao, Junhan; Wang, Yongqiang; Geng, Jianghai; Cheng, Shuyi; Lü, Fangcheng

    2014-08-01

    Fast and accurate positioning of partial discharge (PD) sources in transformer oil is very important for the safe, stable operation of power systems because it allows timely elimination of insulation faults. There is usually more than one PD source once an insulation fault occurs in the transformer oil. This study, which has both theoretical and practical significance, proposes a method of identifying multiple PD sources in the transformer oil. The method combines the two-sided correlation transformation algorithm in the broadband signal focusing and the modified Gerschgorin disk estimator. The method of classification of multiple signals is used to determine the directions of arrival of signals from multiple PD sources. The ultrasonic array positioning method is based on the multi-platform direction finding and the global optimization searching. Both the 4 × 4 square planar ultrasonic sensor array and the ultrasonic array detection platform are built to test the method of identifying and positioning multiple PD sources. The obtained results verify the validity and the engineering practicability of this method.

  20. Nonuniform fast Fourier transform method for numerical diffraction simulation on tilted planes.

    PubMed

    Xiao, Yu; Tang, Xiahui; Qin, Yingxiong; Peng, Hao; Wang, Wei; Zhong, Lijing

    2016-10-01

    The method, based on the rotation of the angular spectrum in the frequency domain, is generally used for the diffraction simulation between the tilted planes. Due to the rotation of the angular spectrum, the interval between the sampling points in the Fourier domain is not even. For the conventional fast Fourier transform (FFT)-based methods, a spectrum interpolation is needed to get the approximate sampling value on the equidistant sampling points. However, due to the numerical error caused by the spectrum interpolation, the calculation accuracy degrades very quickly as the rotation angle increases. Here, the diffraction propagation between the tilted planes is transformed into a problem about the discrete Fourier transform on the uneven sampling points, which can be evaluated effectively and precisely through the nonuniform fast Fourier transform method (NUFFT). The most important advantage of this method is that the conventional spectrum interpolation is avoided and the high calculation accuracy can be guaranteed for different rotation angles, even when the rotation angle is close to π/2. Also, its calculation efficiency is comparable with that of the conventional FFT-based methods. Numerical examples as well as a discussion about the calculation accuracy and the sampling method are presented.

  1. Research of on-line monitoring method for insulation condition of power transformer bushing

    NASA Astrophysics Data System (ADS)

    Xia, Jiuyun; Qian, Zheng; Yu, Hao; Yao, Junda

    2016-01-01

    The power transformer is the key equipment of the power system; its insulation condition will directly influence the security and reliability of the power system. Thus, the on-line monitoring of power transformer is urgently required in order to guarantee the normal operation of the power system. Moreover, the dielectric loss factor is a significant parameter reflecting the condition of transformer bushing, so the on-line measurement of dielectric loss factor is really important. In this paper, the phase-to-phase comparison method is selected as the on-line monitoring method based on the overall analysis and discussion of the existing on-line monitoring methods. At first, the harmonic analysis method is utilized to calculate the dielectric loss of each phase of the three-phase transformer bushing, and then the differences of dielectric loss between every two phases are calculated and analyzed. So the insulation condition of each bushing could be achieved based on the careful analysis of different phase-to-phase dielectric loss. The simulation results of phase-to-phase comparison method are carried out in this paper, and the validity is verified. At last, this method is utilized in an actual equipment of on-line monitoring.

  2. Research on electrocardiogram baseline wandering correction based on wavelet transform, QRS barycenter fitting, and regional method.

    PubMed

    Song, Jinzhong; Yan, Hong; Li, Yanjun; Mu, Kaiyu

    2010-09-01

    Baseline wandering in electrocardiogram (ECG) is one of the biggest interferences in visualization and computerized detection of waveforms (especially ST-segment) based on threshold decision. A new method based on wavelet transform, QRS barycenter fitting and regional method was proposed in this paper. Firstly, wavelet transform as a coarse correction was used to remove the baseline wandering, whose frequency bands were non-overlapping with that of ST-segment. Secondly, QRS barycenter fitting was applied as a detailed correction. The third, the regional method was used to transfer baseline to zero. Finally, the method in this paper was proved to perform better than filtering and function fitting methods in baseline wandering correction after the long-term ST database (LTST) verification. In addition, the proposed method is simple and easy to carry out, and in current use.

  3. An effective numerical method to solve a class of nonlinear singular boundary value problems using improved differential transform method.

    PubMed

    Xie, Lie-Jun; Zhou, Cai-Lian; Xu, Song

    2016-01-01

    In this work, an effective numerical method is developed to solve a class of singular boundary value problems arising in various physical models by using the improved differential transform method (IDTM). The IDTM applies the Adomian polynomials to handle the differential transforms of the nonlinearities arising in the given differential equation. The relation between the Adomian polynomials of those nonlinear functions and the coefficients of unknown truncated series solution is given by a simple formula, through which one can easily deduce the approximate solution which takes the form of a convergent series. An upper bound for the estimation of approximate error is presented. Several physical problems are discussed as illustrative examples to testify the validity and applicability of the proposed method. Comparisons are made between the present method and the other existing methods.

  4. Introduction to the Restoration of Astrophysical Images by Multiscale Transforms and Bayesian Methods

    NASA Astrophysics Data System (ADS)

    Bijaoui, A.

    2013-03-01

    The image restoration is today an important part of the astrophysical data analysis. The denoising and the deblurring can be efficiently performed using multiscale transforms. The multiresolution analysis constitutes the fundamental pillar for these transforms. The discrete wavelet transform is introduced from the theory of the approximation by translated functions. The continuous wavelet transform carries out a generalization of multiscale representations from translated and dilated wavelets. The à trous algorithm furnishes its discrete redundant transform. The image denoising is first considered without any hypothesis on the signal distribution, on the basis of the a contrario detection. Different softening functions are introduced. The introduction of a regularization constraint may improve the results. The application of Bayesian methods leads to an automated adaptation of the softening function to the signal distribution. The MAP principle leads to the basis pursuit, a sparse decomposition on redundant dictionaries. Nevertheless the posterior expectation minimizes, scale per scale, the quadratic error. The proposed deconvolution algorithm is based on a coupling of the wavelet denoising with an iterative inversion algorithm. The different methods are illustrated by numerical experiments on a simulated image similar to images of the deep sky. A white Gaussian stationary noise was added with three levels. In the conclusion different important connected problems are tackled.

  5. Comparisons between real and complex Gauss wavelet transform methods of three-dimensional shape reconstruction

    NASA Astrophysics Data System (ADS)

    Xu, Luopeng; Dan, Youquan; Wang, Qingyuan

    2015-10-01

    The continuous wavelet transform (CWT) introduces an expandable spatial and frequency window which can overcome the inferiority of localization characteristic in Fourier transform and windowed Fourier transform. The CWT method is widely applied in the non-stationary signal analysis field including optical 3D shape reconstruction with remarkable performance. In optical 3D surface measurement, the performance of CWT for optical fringe pattern phase reconstruction usually depends on the choice of wavelet function. A large kind of wavelet functions of CWT, such as Mexican Hat wavelet, Morlet wavelet, DOG wavelet, Gabor wavelet and so on, can be generated from Gauss wavelet function. However, so far, application of the Gauss wavelet transform (GWT) method (i.e. CWT with Gauss wavelet function) in optical profilometry is few reported. In this paper, the method using GWT for optical fringe pattern phase reconstruction is presented first and the comparisons between real and complex GWT methods are discussed in detail. The examples of numerical simulations are also given and analyzed. The results show that both the real GWT method along with a Hilbert transform and the complex GWT method can realize three-dimensional surface reconstruction; and the performance of reconstruction generally depends on the frequency domain appearance of Gauss wavelet functions. For the case of optical fringe pattern of large phase variation with position, the performance of real GWT is better than that of complex one due to complex Gauss series wavelets existing frequency sidelobes. Finally, the experiments are carried out and the experimental results agree well with our theoretical analysis.

  6. An improving fringe analysis method based on the accuracy of S-transform profilometry

    NASA Astrophysics Data System (ADS)

    Shen, Qiuju; Chen, Wenjing; Zhong, Min; Su, Xianyu

    2014-07-01

    The S transform, as a simple and popular technique for spacetime-frequency analysis, has been introduced in optical three-dimensional surface shape measurement in recent years. Based on the S transform, S transform “ridge” method (STR) and S transform filtering method (STF) have been proposed to extract the phase information from the single deformed fringe pattern. This paper focuses on studying the STR in fringe pattern analysis. In previous researches about the STR, a linear constraint, which assumed that the phase was locally expressed as first-order Taylor expansion with respect to x and y directions, was implicitly added. Actually at least the second-order partial derivatives of the phase in each local area should be taken into account because they are related to the local curvature of the height distribution of the tested object. Therefore, the traditional STR has larger phase measuring errors in those areas with the rapid height variation on the tested object. This paper proposes an improved STR method, in which the phase is approximately expressed as a quadric in each local area. The phase extraction formula based on the quadric is derived, and the phase correction is carried out as well. Both the simulations and the experiments verify that a more accurate phase map can be obtained by the improved method compared with that by the traditional STR, especially in the areas where height variation is steep.

  7. The unified transform method for the Sasa–Satsuma equation on the half-line

    PubMed Central

    Xu, Jian; Fan, Engui

    2013-01-01

    We implement the unified transform method to the initial-boundary value (IBV) problem of the Sasa–Satsuma equation on the half line. In addition to presenting the basic Riemann–Hilbert formalism, which linearizes this IBV problem, we also analyse the associated general Dirichlet to Neumann map using the so-called global relation. PMID:24204181

  8. A method of image compression based on lifting wavelet transform and modified SPIHT

    NASA Astrophysics Data System (ADS)

    Lv, Shiliang; Wang, Xiaoqian; Liu, Jinguo

    2016-11-01

    In order to improve the efficiency of remote sensing image data storage and transmission we present a method of the image compression based on lifting scheme and modified SPIHT(set partitioning in hierarchical trees) by the design of FPGA program, which realized to improve SPIHT and enhance the wavelet transform image compression. The lifting Discrete Wavelet Transform (DWT) architecture has been selected for exploiting the correlation among the image pixels. In addition, we provide a study on what storage elements are required for the wavelet coefficients. We present lena's image using the 3/5 lifting scheme.

  9. Lane detection and tracking based on improved Hough transform and least-squares method

    NASA Astrophysics Data System (ADS)

    Sun, Peng; Chen, Hui

    2014-11-01

    Lane detection and tracking play important roles in lane departure warning system (LDWS). In order to improve the real-time performance and obtain better lane detection results, an improved algorithm of lane detection and tracking based on combination of improved Hough transform and least-squares fitting method is proposed in this paper. In the image pre-processing stage, firstly a multi-gradient Sobel operator is used to obtain the edge map of road images, secondly adaptive Otsu algorithm is used to obtain binary image, and in order to meet the precision requirements of single pixel, fast parallel thinning algorithm is used to get the skeleton map of binary image. And then, lane lines are initially detected by using polar angle constraint Hough transform, which has narrowed the scope of searching. At last, during the tracking phase, based on the detection result of the previous image frame, a dynamic region of interest (ROI) is set up, and within the predicted dynamic ROI, least-squares fitting method is used to fit the lane line, which has greatly reduced the algorithm calculation. And also a failure judgment module is added in this paper to improve the detection reliability. When the least-squares fitting method is failed, the polar angle constraint Hough transform is restarted for initial detection, which has achieved a coordination of Hough transform and least-squares fitting method. The algorithm in this paper takes into account the robustness of Hough transform and the real-time performance of least-squares fitting method, and sets up a dynamic ROI for lane detection. Experimental results show that it has a good performance of lane recognition, and the average time to complete the preprocessing and lane recognition of one road map is less than 25ms, which has proved that the algorithm has good real-time performance and strong robustness.

  10. Study on identifying deciduous forest by the method of feature space transformation

    NASA Astrophysics Data System (ADS)

    Zhang, Xuexia; Wu, Pengfei

    2009-10-01

    The thematic remotely sensed information extraction is always one of puzzling nuts which the remote sensing science faces, so many remote sensing scientists devotes diligently to this domain research. The methods of thematic information extraction include two kinds of the visual interpretation and the computer interpretation, the developing direction of which is intellectualization and comprehensive modularization. The paper tries to develop the intelligent extraction method of feature space transformation for the deciduous forest thematic information extraction in Changping district of Beijing city. The whole Chinese-Brazil resources satellite images received in 2005 are used to extract the deciduous forest coverage area by feature space transformation method and linear spectral decomposing method, and the result from remote sensing is similar to woodland resource census data by Chinese forestry bureau in 2004.

  11. [An automatic peak detection method for LIBS spectrum based on continuous wavelet transform].

    PubMed

    Chen, Peng-Fei; Tian, Di; Qiao, Shu-Jun; Yang, Guang

    2014-07-01

    Spectrum peak detection in the laser-induced breakdown spectroscopy (LIBS) is an essential step, but the presence of background and noise seriously disturb the accuracy of peak position. The present paper proposed a method applied to automatic peak detection for LIBS spectrum in order to enhance the ability of overlapping peaks searching and adaptivity. We introduced the ridge peak detection method based on continuous wavelet transform to LIBS, and discussed the choice of the mother wavelet and optimized the scale factor and the shift factor. This method also improved the ridge peak detection method with a correcting ridge method. The experimental results show that compared with other peak detection methods (the direct comparison method, derivative method and ridge peak search method), our method had a significant advantage on the ability to distinguish overlapping peaks and the precision of peak detection, and could be be applied to data processing in LIBS.

  12. The Application of Continuous Wavelet Transform Based Foreground Subtraction Method in 21 cm Sky Surveys

    NASA Astrophysics Data System (ADS)

    Gu, Junhua; Xu, Haiguang; Wang, Jingying; An, Tao; Chen, Wen

    2013-08-01

    We propose a continuous wavelet transform based non-parametric foreground subtraction method for the detection of redshifted 21 cm signal from the epoch of reionization. This method works based on the assumption that the foreground spectra are smooth in frequency domain, while the 21 cm signal spectrum is full of saw-tooth-like structures, thus their characteristic scales are significantly different. We can distinguish them in the wavelet coefficient space easily and perform the foreground subtraction. Compared with the traditional spectral fitting based method, our method is more tolerant to complex foregrounds. Furthermore, we also find that when the instrument has uncorrected response error, our method can also work significantly better than the spectral fitting based method. Our method can obtain similar results with the Wp smoothing method, which is also a non-parametric method, but our method consumes much less computing time.

  13. Automatic active space selection for the similarity transformed equations of motion coupled cluster method

    NASA Astrophysics Data System (ADS)

    Dutta, Achintya Kumar; Nooijen, Marcel; Neese, Frank; Izsák, Róbert

    2017-02-01

    An efficient scheme for the automatic selection of an active space for similarity transformed equations of motion (STEOM) coupled cluster method is proposed. It relies on state averaged configuration interaction singles (CIS) natural orbitals and makes it possible to use STEOM as a black box method. The performance of the new scheme is tested for singlet and triplet valence, charge transfer, and Rydberg excited states.

  14. A complex guided spectral transform Lanczos method for studying quantum resonance states

    DOE PAGES

    Yu, Hua-Gen

    2014-12-28

    A complex guided spectral transform Lanczos (cGSTL) algorithm is proposed to compute both bound and resonance states including energies, widths and wavefunctions. The algorithm comprises of two layers of complex-symmetric Lanczos iterations. A short inner layer iteration produces a set of complex formally orthogonal Lanczos (cFOL) polynomials. They are used to span the guided spectral transform function determined by a retarded Green operator. An outer layer iteration is then carried out with the transform function to compute the eigen-pairs of the system. The guided spectral transform function is designed to have the same wavefunctions as the eigenstates of the originalmore » Hamiltonian in the spectral range of interest. Therefore the energies and/or widths of bound or resonance states can be easily computed with their wavefunctions or by using a root-searching method from the guided spectral transform surface. The new cGSTL algorithm is applied to bound and resonance states of HO₂, and compared to previous calculations.« less

  15. A complex guided spectral transform Lanczos method for studying quantum resonance states

    SciTech Connect

    Yu, Hua-Gen

    2014-12-28

    A complex guided spectral transform Lanczos (cGSTL) algorithm is proposed to compute both bound and resonance states including energies, widths and wavefunctions. The algorithm comprises of two layers of complex-symmetric Lanczos iterations. A short inner layer iteration produces a set of complex formally orthogonal Lanczos (cFOL) polynomials. They are used to span the guided spectral transform function determined by a retarded Green operator. An outer layer iteration is then carried out with the transform function to compute the eigen-pairs of the system. The guided spectral transform function is designed to have the same wavefunctions as the eigenstates of the original Hamiltonian in the spectral range of interest. Therefore the energies and/or widths of bound or resonance states can be easily computed with their wavefunctions or by using a root-searching method from the guided spectral transform surface. The new cGSTL algorithm is applied to bound and resonance states of HO₂, and compared to previous calculations.

  16. Transformation-cost time-series method for analyzing irregularly sampled data.

    PubMed

    Ozken, Ibrahim; Eroglu, Deniz; Stemler, Thomas; Marwan, Norbert; Bagci, G Baris; Kurths, Jürgen

    2015-06-01

    Irregular sampling of data sets is one of the challenges often encountered in time-series analysis, since traditional methods cannot be applied and the frequently used interpolation approach can corrupt the data and bias the subsequence analysis. Here we present the TrAnsformation-Cost Time-Series (TACTS) method, which allows us to analyze irregularly sampled data sets without degenerating the quality of the data set. Instead of using interpolation we consider time-series segments and determine how close they are to each other by determining the cost needed to transform one segment into the following one. Using a limited set of operations-with associated costs-to transform the time series segments, we determine a new time series, that is our transformation-cost time series. This cost time series is regularly sampled and can be analyzed using standard methods. While our main interest is the analysis of paleoclimate data, we develop our method using numerical examples like the logistic map and the Rössler oscillator. The numerical data allows us to test the stability of our method against noise and for different irregular samplings. In addition we provide guidance on how to choose the associated costs based on the time series at hand. The usefulness of the TACTS method is demonstrated using speleothem data from the Secret Cave in Borneo that is a good proxy for paleoclimatic variability in the monsoon activity around the maritime continent.

  17. Fast-Fourier-transform based numerical integration method for the Rayleigh-Sommerfeld diffraction formula

    NASA Astrophysics Data System (ADS)

    Shen, Fabin; Wang, Anbo

    2006-02-01

    The numerical calculation of the Rayleigh-Sommerfeld diffraction integral is investigated. The implementation of a fast-Fourier-transform (FFT) based direct integration (FFT-DI) method is presented, and Simpson's rule is used to improve the calculation accuracy. The sampling interval, the size of the computation window, and their influence on numerical accuracy and on computational complexity are discussed for the FFT-DI and the FFT-based angular spectrum (FFT-AS) methods. The performance of the FFT-DI method is verified by numerical simulation and compared with that of the FFT-AS method.

  18. A method extracting solar cell parameters from spectral response by inverse laplace transform

    NASA Astrophysics Data System (ADS)

    Tuominen, E.; Acerbis, M.; Hovinen, A.; Siirtola, T.; Sinkkonen, J.

    1997-01-01

    A mathematical method to interpret spectral responses measured from solar cells has been developed. Taking an inverse Laplace transform from the spectral response of a solar cell the spatial dependent collection efficiency of the cell can be obtained. Several important material parameters of the solar cell can be extracted from this function. Applying this method the properties of the solar cell can be investigated without applying characterization methods to the cell itself. We have applied the method both to simulated solar cells andto real solar cells.

  19. Agrobacterium tumefaciens-mediated transformation of the entomopathogenic fungus Nomuraea rileyi.

    PubMed

    Shao, Changwen; Yin, Youping; Qi, Zhaoran; Li, Ren; Song, Zhangyong; Li, Yan; Wang, Zhongkang

    2015-10-01

    An Agrobacterium-mediated genetic transformation system for the entomopathogenic fungus Nomuraea rileyi was established. Three binary T-DNA vectors, pPZP-Hph, pPZP-Hph-RNAi and pPZP-Hph-DsRed2, were constructed. The trpc promoter from Aspergillus nidulans was used as the cis-regulatory element to drive the expression of hygromycin phosphotransferase (hph) gene and DsRed2, which conferred the hygromycin B (Hyg B) resistance and red fluorescence visualization, respectively. The blastospores and conidia were used as the recipients. The blastospores' transformation efficiency reached ∼20-40 transformants per 10(6) blastospores, whereas the conidia were not transformed. Based on an analysis of five generations of subcultures, PCR and Southern blotting assays, the Ptrpc-hph cassette had integrated into the genomes of all transformants, which contained single copy of the hph gene and showed mitotic stability. Abundant altered morphologic phenotypes in colonies, blastospores and hyphae formations were observed in the arbitrary insertional mutants of N. rileyi, which made it possible to study the relationships between the functions and the interrupted genes over the whole genome. The transformation protocol will promote the functional characterization of genes, and the construction of genetically engineered strains of this important entomopathogenic fungus, and potentially of other similar fungal pathogens.

  20. New approach application of data transformation in mean centering of ratio spectra method

    NASA Astrophysics Data System (ADS)

    Issa, Mahmoud M.; Nejem, R.'afat M.; Van Staden, Raluca Ioana Stefan; Aboul-Enein, Hassan Y.

    2015-05-01

    Most of mean centering (MCR) methods are designed to be used with data sets whose values have a normal or nearly normal distribution. The errors associated with the values are also assumed to be independent and random. If the data are skewed, the results obtained may be doubtful. Most of the time, it was assumed a normal distribution and if a confidence interval includes a negative value, it was cut off at zero. However, it is possible to transform the data so that at least an approximately normal distribution is attained. Taking the logarithm of each data point is one transformation frequently used. As a result, the geometric mean is deliberated a better measure of central tendency than the arithmetic mean. The developed MCR method using the geometric mean has been successfully applied to the analysis of a ternary mixture of aspirin (ASP), atorvastatin (ATOR) and clopidogrel (CLOP) as a model. The results obtained were statistically compared with reported HPLC method.

  1. The performance of methods based on the fractional Fourier transform for detecting marine mammal vocalizations.

    PubMed

    Locke, Jonathan; White, Paul R

    2011-10-01

    The analysis of cetacean vocalizations is considered using Fourier-based techniques that employ chirp functions in their decomposition. In particular, the paper considers a short-time methods based on the fractional Fourier transform for detecting frequency modulated narrow-band signals, such as dolphin whistles, and compares this to the classical short-time Fourier methods. The fractional Fourier technique explored computes transforms associated with a range of chirp rates and automatically selects the rate for the final analysis. This avoids the need for prior knowledge of signal's chirp rate. An analysis is presented that details the performance of both methods as signal detectors and allows one to determine their detection thresholds. These thresholds are then used to measure the detectability of synthetic signals. This principle is then extended to measure performance on a set of recordings of narrow-band vocalizations from a range of cetacean species.

  2. Solution of nonlinear higher-index Hessenberg DAEs by Adomian polynomials and differential transform method.

    PubMed

    Benhammouda, Brahim

    2015-01-01

    The solution of higher-index Hessenberg differential-algebraic equations (DAEs) is of great importance since this type of DAEs often arises in applications. Higher-index DAEs are known to be numerically and analytically difficult to solve. In this paper, we present a new analytical method for the solution of two classes of higher-index Hessenberg DAEs. The method is based on Adomian polynomials and the differential transform method (DTM). First, the DTM is applied to the DAE where the differential transforms of nonlinear terms are calculated using Adomian polynomials. Then, based on the index condition, the resulting recursion system is transformed into a nonsingular linear algebraic system. This system is then solved to obtain the coefficients of the power series solution. The main advantage of the proposed technique is that it does not require an index reduction nor a linearization. Two test problems are solved to demonstrate the effectiveness of the method. In addition, to extend the domain of convergence of the approximate series solution, we propose a post-treatment with Laplace-Padé resummation method.

  3. Are we on the brink of a major transformation of clinical method?

    PubMed Central

    McWhinney, I R

    1986-01-01

    The traditional clinical method, which has served medicine well for over 100 years, had its origins in the integration of physical examination with morbid anatomy in early 19th-century France. Now this method is showing signs of failing to meet some contemporary needs. In particular, there is no means for understanding the inner experience of patients. Previous models of a transformed method have not grappled with the problem of validation. Data on the inner experience of patients are not open to empiric validation in the same way as clinical data. The process of understanding the meaning of an illness is not, therefore, scientific in the conventional sense. There are, none the less, rigorous methods for validating the results of this form of inquiry, notably those of phenomenology. A transformed method should aim to understand the meaning of an illness for the patient as well as provide a clinical diagnosis. The transformation will require a change in the epistemology of medicine and an educational process that encourages reflection and growth of self-knowledge. PMID:3756718

  4. Analysis of comparative efficiencies of different transformation methods of E. coli using two common plasmid vectors.

    PubMed

    Roychoudhury, Aryadeep; Basu, Supratim; Sengupta, Dibyendu N

    2009-10-01

    The efficiencies of different transformation methods of E. coli DH5Qalpha train, induced by several cations like Mg2+, Mn2+ Rb+ and especially Ca2+, with or without polyethylene glycol (PEG) and dimethyl sulfoxide (DMSO) were compared using the two commonly used plasmid vectors pCAMBIA1201 and pBI121. The widely used calcium chloride (CaCl2) method appeared to be the most efficient procedure, while rubidium chloride (RbCl) method was the least effective. The improvements in the classical CaCl2 method were found to further augment the transformation efficiency (TR)E for both the vectors like repeated alternate cycles of heat shock, followed by immediate cold, at least up to the third cycle; replacement of the heat shock step by a single microwave pulse and even more by double microwave treatment and administration of combined heat shock-microwave treatments. The pre-treatment of CaCl2-competent cells with 5% (v/v) ethanol, accompanied by single heat shock also triggered the (TR)E, which was further enhanced, when combined heat shock-microwave was applied. The minor alterations or improved approaches in CaCl2 method suggested in the present study may thus find use in more efficient E. coli transformation.

  5. Method to eliminate flux linkage DC component in load transformer for static transfer switch.

    PubMed

    He, Yu; Mao, Chengxiong; Lu, Jiming; Wang, Dan; Tian, Bing

    2014-01-01

    Many industrial and commercial sensitive loads are subject to the voltage sags and interruptions. The static transfer switch (STS) based on the thyristors is applied to improve the power quality and reliability. However, the transfer will result in severe inrush current in the load transformer, because of the DC component in the magnetic flux generated in the transfer process. The inrush current which is always 2 ~ 30 p.u. can cause the disoperation of relay protective devices and bring potential damage to the transformer. The way to eliminate the DC component is to transfer the related phases when the residual flux linkage of the load transformer and the prospective flux linkage of the alternate source are equal. This paper analyzes how the flux linkage of each winding in the load transformer changes in the transfer process. Based on the residual flux linkage when the preferred source is completely disconnected, the method to calculate the proper time point to close each phase of the alternate source is developed. Simulation and laboratory experiments results are presented to show the effectiveness of the transfer method.

  6. Method to Eliminate Flux Linkage DC Component in Load Transformer for Static Transfer Switch

    PubMed Central

    2014-01-01

    Many industrial and commercial sensitive loads are subject to the voltage sags and interruptions. The static transfer switch (STS) based on the thyristors is applied to improve the power quality and reliability. However, the transfer will result in severe inrush current in the load transformer, because of the DC component in the magnetic flux generated in the transfer process. The inrush current which is always 2~30 p.u. can cause the disoperation of relay protective devices and bring potential damage to the transformer. The way to eliminate the DC component is to transfer the related phases when the residual flux linkage of the load transformer and the prospective flux linkage of the alternate source are equal. This paper analyzes how the flux linkage of each winding in the load transformer changes in the transfer process. Based on the residual flux linkage when the preferred source is completely disconnected, the method to calculate the proper time point to close each phase of the alternate source is developed. Simulation and laboratory experiments results are presented to show the effectiveness of the transfer method. PMID:25133255

  7. An automatic recognition method of pointer instrument based on improved Hough transform

    NASA Astrophysics Data System (ADS)

    Xu, Li; Fang, Tian; Gao, Xiaoyu

    2015-10-01

    For the automatic recognition of pointer instrument, the method for the automatic recognition of pointer instrument based on improved Hough Transform was proposed in this paper. The automatic recognition of pointer instrument is applied to all kinds of lighting conditions, but the accuracy of it binaryzation will be influenced when the light is too strong or too dark. Therefore, the improved Ostu method was suggested to realize recognition for adaptive thresholding of pointer instrument under all kinds of lighting conditions. On the basis of dial image characteristics, Otsu method is used to get the value of maximum between-cluster variance and initial threshold than analyze its maximum between-cluster variance value to determine the light and shade of the image. When the images are too bright or too dark, the smaller pixels should be given up and then calculate the initial threshold by Otsu method again and again until the best binaryzation image was obtained. Hence, transform the pointer straight line of the binaryzation image to Hough parameter space through improved Hough Transform to determine the position of the pointer straight line by searching the maximum value of arrays of the same angle. Finally, according to angle method, the pointer reading was obtained by the linear relationship for the initial scale and angle of the pointer instrument. Results show that the improved Otsu method make pointer instrument possible to obtained the accuracy binaryzation image even though the light is too bright or too dark , which improves the adaptability of pointer instrument to automatic recognize the light under different conditions. For the pressure gauges with range of 60MPa, the relative error identification reached to 0.005 when use the improved Hough Transform Algorithm.

  8. A new stationary gridline artifact suppression method based on the 2D discrete wavelet transform

    PubMed Central

    Tang, Hui; Tong, Dan; Bao, Xudong; Dillenseger, Jean-Louis

    2015-01-01

    Purpose In digital X-ray radiography, an anti-scatter grid is inserted between the patient and the image receptor to reduce scattered radiation. If the anti-scatter grid is used in a stationary way, gridline artifacts will appear in the final image. In most of the gridline removal image processing methods, the useful information with spatial frequencies close to that of the gridline is usually lost or degraded. In this study, a new stationary gridline suppression method is designed to preserve more of the useful information. Methods The method is as follows. The input image is first recursively decomposed into several smaller sub-images using a multi-scale 2D discrete wavelet transform (DWT). The decomposition process stops when the gridline signal is found to be greater than a threshold in one or several of these sub-images using a gridline detection module. An automatic Gaussian band-stop filter is then applied to the detected sub-images to remove the gridline signal. Finally, the restored image is achieved using the corresponding 2D inverse discrete wavelet transform (IDWT). Results The processed images show that the proposed method can remove the gridline signal efficiently while maintaining the image details. The spectra of a 1-dimensional Fourier transform of the processed images demonstrate that, compared with some existing gridline removal methods, the proposed method has better information preservation after the removal of the gridline artifacts. Additionally, the performance speed is relatively high. Conclusions The experimental results demonstrate the efficiency of the proposed method. Compared with some existing gridline removal methods, the proposed method can preserve more information within an acceptable execution time. PMID:25832061

  9. A new stationary gridline artifact suppression method based on the 2D discrete wavelet transform

    SciTech Connect

    Tang, Hui; Tong, Dan; Dong Bao, Xu; Dillenseger, Jean-Louis

    2015-04-15

    Purpose: In digital x-ray radiography, an antiscatter grid is inserted between the patient and the image receptor to reduce scattered radiation. If the antiscatter grid is used in a stationary way, gridline artifacts will appear in the final image. In most of the gridline removal image processing methods, the useful information with spatial frequencies close to that of the gridline is usually lost or degraded. In this study, a new stationary gridline suppression method is designed to preserve more of the useful information. Methods: The method is as follows. The input image is first recursively decomposed into several smaller subimages using a multiscale 2D discrete wavelet transform. The decomposition process stops when the gridline signal is found to be greater than a threshold in one or several of these subimages using a gridline detection module. An automatic Gaussian band-stop filter is then applied to the detected subimages to remove the gridline signal. Finally, the restored image is achieved using the corresponding 2D inverse discrete wavelet transform. Results: The processed images show that the proposed method can remove the gridline signal efficiently while maintaining the image details. The spectra of a 1D Fourier transform of the processed images demonstrate that, compared with some existing gridline removal methods, the proposed method has better information preservation after the removal of the gridline artifacts. Additionally, the performance speed is relatively high. Conclusions: The experimental results demonstrate the efficiency of the proposed method. Compared with some existing gridline removal methods, the proposed method can preserve more information within an acceptable execution time.

  10. High-capacity method for hiding data in the discrete cosine transform domain

    NASA Astrophysics Data System (ADS)

    Qazanfari, Kazem; Safabakhsh, Reza

    2013-10-01

    Steganography is the art and science of hiding data in different media such as texts, audios, images, and videos. Data hiding techniques are generally divided into two groups: spatial and frequency domain techniques. Spatial domain methods generally have low security and, as a result, are less attractive to researchers. Discrete cosine transform (DCT) is the most common transform domain used in steganography and JPEG compression. Since a large number of the DCT coefficients of JPEG images are zero, the capacity of DCT domain-based steganography methods is not very high. We present a high-capacity method for hiding messages in the DCT domain. We describe the method in two classes where the receiver has and where the receiver does not have the cover image. In each class, we consider three cases for each coefficient. By considering n coefficients, there are 3n different situations. The method embeds ⌊log2 3n⌋ bits in these n coefficients. We show that the maximum reachable capacity by our method is 58% higher than the other general steganography methods. Experimental results show that the histogram-based steganalysis methods cannot detect the stego images produced by the proposed method while the capacity is increased significantly.

  11. Trace projection transformation: a new method for measurement of debris flow surface velocity fields

    NASA Astrophysics Data System (ADS)

    Yan, Yan; Cui, Peng; Guo, Xiaojun; Ge, Yonggang

    2016-12-01

    Spatiotemporal variation of velocity is important for debris flow dynamics. This paper presents a new method, the trace projection transformation, for accurate, non-contact measurement of a debris-flow surface velocity field based on a combination of dense optical flow and perspective projection transformation. The algorithm for interpreting and processing is implemented in C ++ and realized in Visual Studio 2012. The method allows quantitative analysis of flow motion through videos from various angles (camera positioned at the opposite direction of fluid motion). It yields the spatiotemporal distribution of surface velocity field at pixel level and thus provides a quantitative description of the surface processes. The trace projection transformation is superior to conventional measurement methods in that it obtains the full surface velocity field by computing the optical flow of all pixels. The result achieves a 90% accuracy of when comparing with the observed values. As a case study, the method is applied to the quantitative analysis of surface velocity field of a specific debris flow.

  12. Periodic nonlinear Fourier transform for fiber-optic communications, Part I: theory and numerical methods.

    PubMed

    Kamalian, Morteza; Prilepsky, Jaroslaw E; Le, Son Thai; Turitsyn, Sergei K

    2016-08-08

    In this work, we introduce the periodic nonlinear Fourier transform (PNFT) method as an alternative and efficacious tool for compensation of the nonlinear transmission effects in optical fiber links. In the Part I, we introduce the algorithmic platform of the technique, describing in details the direct and inverse PNFT operations, also known as the inverse scattering transform for periodic (in time variable) nonlinear Schrödinger equation (NLSE). We pay a special attention to explaining the potential advantages of the PNFT-based processing over the previously studied nonlinear Fourier transform (NFT) based methods. Further, we elucidate the issue of the numerical PNFT computation: we compare the performance of four known numerical methods applicable for the calculation of nonlinear spectral data (the direct PNFT), in particular, taking the main spectrum (utilized further in Part II for the modulation and transmission) associated with some simple example waveforms as the quality indicator for each method. We show that the Ablowitz-Ladik discretization approach for the direct PNFT provides the best performance in terms of the accuracy and computational time consumption.

  13. Use of the fast Fourier transform method for analyzing linear and equispaced Fizeau fringes.

    PubMed

    Lai, G; Yatagai, T

    1994-09-01

    The Fourier transform method is applied to analyze the initial phase of linear and equispaced Fizeau fringes. We develop an algorithm for high-precision phase measurement by using the Fourier coefficient that corresponds to the spatial frequency of the Fizeau fringes, and we describe methods for determining the fringe carrier frequency. Errors caused by carrier frequency fluctuation and data truncation are studied theoretically and by computer simulation. To demonstrate the method we apply it to the real-time calibration of a piezoelectric transducer mirror in a Twyman-Green interferometer.

  14. A Fusion Method of Gabor Wavelet Transform and Unsupervised Clustering Algorithms for Tissue Edge Detection

    PubMed Central

    Ergen, Burhan

    2014-01-01

    This paper proposes two edge detection methods for medical images by integrating the advantages of Gabor wavelet transform (GWT) and unsupervised clustering algorithms. The GWT is used to enhance the edge information in an image while suppressing noise. Following this, the k-means and Fuzzy c-means (FCM) clustering algorithms are used to convert a gray level image into a binary image. The proposed methods are tested using medical images obtained through Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) devices, and a phantom image. The results prove that the proposed methods are successful for edge detection, even in noisy cases. PMID:24790590

  15. [Decomposition and analysis of the natural source SLF spectrum using curvelet transform method].

    PubMed

    Jiang, Hong-bo; Chen, Chao; Qin, Qi-ming

    2012-02-01

    Because natural source super low frequency (SLF) electromagnetic detection equipment receives wideband multi-source signal, how to decompose the signal to filter out the interference signal was a key factor for the application of natural source SLF electromagnetic detection technology. In the present article, the detection equipment developed by Peking University was used to survey the coal bed methane data in the Qinshui basin, Shanxi province, and the curvelet transform method was employed to decompose those data. The analysis results indicated that the high-frequency information coming from the decomposition is the interference signals mainly generated by lightning in the atmospheric and directly received by the detection equipment, while the low frequency signal mainly contains the target information. So the reconstructed curve based on the low-frequency information was more favorable for the interpretation of the target, compared with the original spectrum curve. But the curvelet transform method could not remove the artificial frequency signal.

  16. Free vibration investigation of nano mass sensor using differential transformation method

    NASA Astrophysics Data System (ADS)

    Zarepour, Misagh; Hosseini, S. Amirhosein; Ghadiri, Majid

    2017-03-01

    In the present study, transverse vibration of nano-cantilever beam with attached mass and two rotational and transverse springs at its end is studied. Resonance frequency of vibrating system is influenced by changing mass particle and stiffness coefficients. Euler-Bernoulli beam theory, nonlocal constitutive equations of Eringen, and Hamilton's principle are used to develop equations of motion. Differential transformation method (DTM) is applied to solve the governing equations of the nanobeam with attached mass particle. Accurate results with minimum mathematical calculation are the advantages of DTM. A detailed parametric study is conducted to investigate the influences of nonlocal parameter. The results can be used in designing of nanoelectromechanical systems. To verify the results, some comparisons are presented between differential transform method results and open literature to show the accuracy of this new approach.

  17. A Novel Medical Freehand Sketch 3D Model Retrieval Method by Dimensionality Reduction and Feature Vector Transformation.

    PubMed

    Jing, Zhang; Sheng, Kang Bao

    2015-01-01

    To assist physicians to quickly find the required 3D model from the mass medical model, we propose a novel retrieval method, called DRFVT, which combines the characteristics of dimensionality reduction (DR) and feature vector transformation (FVT) method. The DR method reduces the dimensionality of feature vector; only the top M low frequency Discrete Fourier Transform coefficients are retained. The FVT method does the transformation of the original feature vector and generates a new feature vector to solve the problem of noise sensitivity. The experiment results demonstrate that the DRFVT method achieves more effective and efficient retrieval results than other proposed methods.

  18. A Novel Medical Freehand Sketch 3D Model Retrieval Method by Dimensionality Reduction and Feature Vector Transformation

    PubMed Central

    Jing, Zhang; Sheng, Kang Bao

    2016-01-01

    To assist physicians to quickly find the required 3D model from the mass medical model, we propose a novel retrieval method, called DRFVT, which combines the characteristics of dimensionality reduction (DR) and feature vector transformation (FVT) method. The DR method reduces the dimensionality of feature vector; only the top M low frequency Discrete Fourier Transform coefficients are retained. The FVT method does the transformation of the original feature vector and generates a new feature vector to solve the problem of noise sensitivity. The experiment results demonstrate that the DRFVT method achieves more effective and efficient retrieval results than other proposed methods. PMID:27293478

  19. [Application of optimized parameters SVM based on photoacoustic spectroscopy method in fault diagnosis of power transformer].

    PubMed

    Zhang, Yu-xin; Cheng, Zhi-feng; Xu, Zheng-ping; Bai, Jing

    2015-01-01

    In order to solve the problems such as complex operation, consumption for the carrier gas and long test period in traditional power transformer fault diagnosis approach based on dissolved gas analysis (DGA), this paper proposes a new method which is detecting 5 types of characteristic gas content in transformer oil such as CH4, C2H2, C2H4, C2H6 and H2 based on photoacoustic Spectroscopy and C2H2/C2H4, CH4/H2, C2H4/C2H6 three-ratios data are calculated. The support vector machine model was constructed using cross validation method under five support vector machine functions and four kernel functions, heuristic algorithms were used in parameter optimization for penalty factor c and g, which to establish the best SVM model for the highest fault diagnosis accuracy and the fast computing speed. Particles swarm optimization and genetic algorithm two types of heuristic algorithms were comparative studied in this paper for accuracy and speed in optimization. The simulation result shows that SVM model composed of C-SVC, RBF kernel functions and genetic algorithm obtain 97. 5% accuracy in test sample set and 98. 333 3% accuracy in train sample set, and genetic algorithm was about two times faster than particles swarm optimization in computing speed. The methods described in this paper has many advantages such as simple operation, non-contact measurement, no consumption for the carrier gas, long test period, high stability and sensitivity, the result shows that the methods described in this paper can instead of the traditional transformer fault diagnosis by gas chromatography and meets the actual project needs in transformer fault diagnosis.

  20. Notes on a method to transform digitized coordinates to geographic coordinates

    USGS Publications Warehouse

    Evenden, Gerald I.

    1991-01-01

    The process of conversion of coordinates (typically carte- sian) obtained from digitizing devices into application de- ned coordinate systems is discussed. Application of a secondary function representing the transformation of the application coordinate system to and from cartesian space will greatly simplify conversion of highly non-linear sys- tems. Although emphasis is given to cartographic conver- sions the method will work with a wide variety of digitizing problems.

  1. Seismic coherent and random noise attenuation using the undecimated discrete wavelet transform method with WDGA technique

    NASA Astrophysics Data System (ADS)

    Goudarzi, Alireza; Riahi, Mohammad Ali

    2012-12-01

    One of the most crucial challenges in seismic data processing is the reduction of the noise in the data or improving the signal-to-noise ratio. In this study, the 1D undecimated discrete wavelet transform (UDWT) has been acquired to attenuate random noise and ground roll. Wavelet domain ground roll analysis (WDGA) is applied to find the ground roll energy in the wavelet domain. The WDGA will be a substitute method for thresholding in seismic data processing. To compare the effectiveness of the WDGA method, we apply the 1D double density discrete wavelet transform (DDDWT) using soft thresholding in the random noise reduction and ground roll attenuation processes. Seismic signals intersect with ground roll in the time and frequency domains. Random noise and ground roll have many undesirable effects on pre-stack seismic data, and result in an inaccurate velocity analysis for NMO correction. In this paper, the UDWT by using the WDGA technique and DDDWT (using the soft thresholding technique) and the regular Fourier based method as f-k transform will be used and compared for seismic denoising.

  2. [A wavelet-transform-based method for the automatic detection of late-type stars].

    PubMed

    Liu, Zhong-tian; Zhao, Rrui-zhen; Zhao, Yong-heng; Wu, Fu-chao

    2005-07-01

    The LAMOST project, the world largest sky survey project, urgently needs an automatic late-type stars detection system. However, to our knowledge, no effective methods for automatic late-type stars detection have been reported in the literature up to now. The present study work is intended to explore possible ways to deal with this issue. Here, by "late-type stars" we mean those stars with strong molecule absorption bands, including oxygen-rich M, L and T type stars and carbon-rich C stars. Based on experimental results, the authors find that after a wavelet transform with 5 scales on the late-type stars spectra, their frequency spectrum of the transformed coefficient on the 5th scale consistently manifests a unimodal distribution, and the energy of frequency spectrum is largely concentrated on a small neighborhood centered around the unique peak. However, for the spectra of other celestial bodies, the corresponding frequency spectrum is of multimodal and the energy of frequency spectrum is dispersible. Based on such a finding, the authors presented a wavelet-transform-based automatic late-type stars detection method. The proposed method is shown by extensive experiments to be practical and of good robustness.

  3. Remote Sensing Image Fusion Method Based on Nonsubsampled Shearlet Transform and Sparse Representation

    NASA Astrophysics Data System (ADS)

    Moonon, Altan-Ulzii; Hu, Jianwen; Li, Shutao

    2015-12-01

    The remote sensing image fusion is an important preprocessing technique in remote sensing image processing. In this paper, a remote sensing image fusion method based on the nonsubsampled shearlet transform (NSST) with sparse representation (SR) is proposed. Firstly, the low resolution multispectral (MS) image is upsampled and color space is transformed from Red-Green-Blue (RGB) to Intensity-Hue-Saturation (IHS). Then, the high resolution panchromatic (PAN) image and intensity component of MS image are decomposed by NSST to high and low frequency coefficients. The low frequency coefficients of PAN and the intensity component are fused by the SR with the learned dictionary. The high frequency coefficients of intensity component and PAN image are fused by local energy based fusion rule. Finally, the fused result is obtained by performing inverse NSST and inverse IHS transform. The experimental results on IKONOS and QuickBird satellites demonstrate that the proposed method provides better spectral quality and superior spatial information in the fused image than other remote sensing image fusion methods both in visual effect and object evaluation.

  4. A comparison and optimization of methods and factors affecting the transformation of Escherichia coli.

    PubMed

    Chan, Weng-Tat; Verma, Chandra S; Lane, David P; Gan, Samuel Ken-En

    2013-12-12

    DNA manipulation routinely requires competent bacteria that can be made using one of numerous methods. To determine the best methods, we compared four commonly used chemical methods (DMSO, MgCl2-CaCl2, CaCl2 and Hanahan's methods) on frequently used Escherichia coli (E. coli) strains: DH5α, XL-1 Blue, SCS110, JM109, TOP10 and BL21-(DE3)-PLysS. Hanahan's method was found to be most effective for DH5α, XL-1 Blue and JM109 strains (P<0.05), whilst the CaCl2 method was best for SCS110, TOP10 and BL21 strains (P<0.05). The use of SOB (super optimal broth) over LB [Luria-Bertani (broth)] growth media was found to enhance the competency of XL-1 Blue (P<0.05), dampened JM109's competency (P<0.05), and had no effect on the other strains (P>0.05). We found no significant differences between using 45 or 90 s heat shock across all the six strains (P>0.05). Through further optimization by means of concentrating the aliquots, we were able to get further increases in transformation efficiencies. Based on the optimized parameters and methods, these common laboratory E. coli strains attained high levels of TrE (transformation efficiency), thus facilitating the production of highly efficient and cost-effective competent bacteria.

  5. Standardization of size, shape and internal structure of spinal cord images: comparison of three transformation methods.

    PubMed

    Fujiki, Yasuhisa; Yokota, Shigefumi; Okada, Yasumasa; Oku, Yoshitaka; Tamura, Yoshiyasu; Ishiguro, Makio; Miwakeichi, Fumikazu

    2013-01-01

    Functional fluorescence imaging has been widely applied to analyze spatio-temporal patterns of cellular dynamics in the brain and spinal cord. However, it is difficult to integrate spatial information obtained from imaging data in specific regions of interest across multiple samples, due to large variability in the size, shape and internal structure of samples. To solve this problem, we attempted to standardize transversely sectioned spinal cord images focusing on the laminar structure in the gray matter. We employed three standardization methods, the affine transformation (AT), the angle-dependent transformation (ADT) and the combination of these two methods (AT+ADT). The ADT is a novel non-linear transformation method developed in this study to adjust an individual image onto the template image in the polar coordinate system. We next compared the accuracy of these three standardization methods. We evaluated two indices, i.e., the spatial distribution of pixels that are not categorized to any layer and the error ratio by the leave-one-out cross validation method. In this study, we used neuron-specific marker (NeuN)-stained histological images of transversely sectioned cervical spinal cord slices (21 images obtained from 4 rats) to create the standard atlas and also to serve for benchmark tests. We found that the AT+ADT outperformed other two methods, though the accuracy of each method varied depending on the layer. This novel image standardization technique would be applicable to optical recording such as voltage-sensitive dye imaging, and will enable statistical evaluations of neural activation across multiple samples.

  6. Genome-wide profiling of genetic variation in Agrobacterium-transformed rice plants*#

    PubMed Central

    Li, Wen-xu; Wu, San-ling; Liu, Yan-hua; Jin, Gu-lei; Zhao, Hai-jun; Fan, Long-jiang; Shu, Qing-yao

    2016-01-01

    Agrobacterium-mediated transformation has been widely used in producing transgenic plants, and was recently used to generate “transgene-clean” targeted genomic modifications coupled with the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas9) system. Although tremendous variation in morphological and agronomic traits, such as plant height, seed fertility, and grain size, was observed in transgenic plants, the underlying mechanisms are not yet well understood, and the types and frequency of genetic variation in transformed plants have not been fully disclosed. To reveal the genome-wide variation in transformed plants, we sequenced the genomes of five independent T0 rice plants using next-generation sequencing (NGS) techniques. Bioinformatics analyses followed by experimental validation revealed the following: (1) in addition to transfer-DNA (T-DNA) insertions, three transformed plants carried heritable plasmid backbone DNA of variable sizes (855–5216 bp) and in different configurations with the T-DNA insertions (linked or apart); (2) each transgenic plant contained an estimated 338–1774 independent genetic variations (single nucleotide variations (SNVs) or small insertion/deletions); and (3) 2–6 new Tos17 insertions were detected in each transformed plant, but no other transposable elements or bacterial genomic DNA. PMID:27921404

  7. Method of Global Transformation and its Role in Turning Point Problems

    NASA Astrophysics Data System (ADS)

    McArthur, Raymond Peter

    In this monograph, we introduce an asymptotic technique which can be used to obtain uniform asymptotic solutions of certain types of second-order, linear ordinary differential equations which may arise in wave propagation problems in non-homogeneous media. Of particular interest are those physical phenomenon which lead to simple and non-simple turning point problems where the nature of the fundamental solution undergoes a sudden metamorphosis of character at the turning point. We begin by giving a few examples which under certain conditions admit turning point behavior. The general theory of the method of global transformation is then introduced. After giving a preview of the method, a historical account of the idea of comparison equation is given which systematically leads to our generalization of the concept. Next, we transform the original problem to an appropriate comparison equation using new dependent and independent variables. This leads to a non-linear differential equation in the transformed coordinate. This non-linear equation has the property that its solution can be determined in the form of uniformly convergent series. We take advantage of this unique property and develop an iterative technique to obtain the solution of the original problem. In applying our iterative technique, the zero -order solution so obtained reproduces exactly the dominant solutions as given by WKBJ or method of stationary phase. We then obtain the higher-order solutions of the non-linear equation which yield new transformations. Use of these transformations in conjunction with the comparison equation give us higher-order correction terms in the asymptotic series solution of the original problem. These higher -order terms correspond to the higher-order terms which one would obtain by using the method of steepest descent. After giving an account of the well-known connection problem using our zero-order solution of the Airy equation, we conclude by giving an illustrative example of

  8. An Efficient Method to Recognize Geologic Structures by Analyzing the Pressure Data with Wavelet Transform

    NASA Astrophysics Data System (ADS)

    Zhou, H.; Srinivasan, S.; Li, L.; Bryant, S. L.

    2013-12-01

    Uncertainty in prediction of flow performance stems from the uncertainty in model parameters such as conductivity, porosity etc., to a large extent, while the characterization of the model parameters is demanding due to the inherent heterogeneity of geologic structures. Inverse modeling approaches attempt to identify the unknown model structures and corresponding parameters by integrating observation data. Several inverse methods have been proposed in the literature ranging from trial-and-error methods to advanced ensemble Kalman filter assimilation, including those that use multiple point statistics to characterize complex geologic structures. However, these methods are hindered by the huge amount of data accumulated with time, for instance, the pressure data are recorded at very fine time intervals from the very early stage of bore hole drilling to mature production period. Assimilation of such large amount of data can be a computational burden to the inverse methods. The object of this work is to propose a computationally efficient approach to analyze the long observation records in order to recognize the subsurface structures, especially flow connectivity which plays a critical role in transport prediction. Wavelet transform is found to be a powerful technique that transforms data into different components and analyzes each component at corresponding scale. By analyzing the components transformed we relate the characteristics of the heterogeneity to signature in the production/injection records. Combining components at different scales we are able to recognize connectivity between wells, and thereby identify complex structure in aquifers. The method is demonstrated in a synthetic example where CO2 is injected into a deep saline aquifer for sequestration. The method is computationally efficient since it involves no iterative forward simulation or sensitivity matrix computation. Once the important episodes have been identified in the dynamic data, inverse

  9. Laplace transform homotopy perturbation method for the approximation of variational problems.

    PubMed

    Filobello-Nino, U; Vazquez-Leal, H; Rashidi, M M; Sedighi, H M; Perez-Sesma, A; Sandoval-Hernandez, M; Sarmiento-Reyes, A; Contreras-Hernandez, A D; Pereyra-Diaz, D; Hoyos-Reyes, C; Jimenez-Fernandez, V M; Huerta-Chua, J; Castro-Gonzalez, F; Laguna-Camacho, J R

    2016-01-01

    This article proposes the application of Laplace Transform-Homotopy Perturbation Method and some of its modifications in order to find analytical approximate solutions for the linear and nonlinear differential equations which arise from some variational problems. As case study we will solve four ordinary differential equations, and we will show that the proposed solutions have good accuracy, even we will obtain an exact solution. In the sequel, we will see that the square residual error for the approximate solutions, belongs to the interval [0.001918936920, 0.06334882582], which confirms the accuracy of the proposed methods, taking into account the complexity and difficulty of variational problems.

  10. Laplace transform method for linear sequential Riemann Liouville and Caputo fractional differential equations

    NASA Astrophysics Data System (ADS)

    Vatsala, Aghalaya S.; Sowmya, M.

    2017-01-01

    Study of nonlinear sequential fractional differential equations of Riemann-Lioville type and Caputo type initial value problem are very useful in applications. In order to develop any iterative methods to solve the nonlinear problems, we need to solve the corresponding linear problem. In this work, we develop Laplace transform method to solve the linear sequential Riemann-Liouville fractional differential equations as well as linear sequential Caputo fractional differential equations of order nq which is sequential of order q. Also, nq is chosen such that (n-1) < nq < n. All our results yield the integer results as a special case when q tends to 1.

  11. Preliminary Method for Direct Quantification of Colistin Methanesulfonate by Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy

    PubMed Central

    Niece, Krista L.

    2015-01-01

    Colistin use has increased in response to the advent of infections caused by multidrug-resistant organisms. It is administered parenterally as an inactive prodrug, colistin methanesulfonate (CMS). Various formulations of CMS and labeling conventions can lead to confusion about colistin dosing, and questions remain about the pharmacokinetics of CMS. Since CMS does not have strong UV absorbance, current methods employ a laborious process of chemical conversion to colistin followed by precolumn derivatization to detect formed colistin by high-performance liquid chromatography. Here, we report a method for direct quantification of colistin methanesulfonate by attenuated total reflectance Fourier transform infrared spectroscopy (ATR FTIR). PMID:26124160

  12. Instantaneous rest-frame transformation method for temporally induced pair creation

    NASA Astrophysics Data System (ADS)

    Lv, Q. Z.; Vikartofsky, A.; Norris, S.; Li, Y. J.; Wagner, R.; Su, Q.; Grobe, R.

    2014-04-01

    We introduce a computational method to determine the rate of the electron-positron pair creation triggered by a time-dependent subcritical external field. It is based on constructing a Lorentz transformation to an instantaneous rest frame, for which the pair-creation rate can be determined by standard techniques. We will discuss the accuracy and efficiency of this method by comparing its predictions with exact time-dependent quantum field theoretical solutions to the Dirac and Klein-Gordon equations for various space-time dependent external fields.

  13. Dechlorination of polychlorinated biphenyls in industrial transformer oil by radiolytic and photolytic methods.

    PubMed

    Jones, Cynthia G; Silverman, Joseph; Al-Sheikhly, Mohamad; Neta, Pedatsur; Poster, Dianne L

    2003-12-15

    Used electrical transformer oils containing low or high concentrations of polychlorinated biphenyls (PCBs) were treated using electron, gamma, and ultraviolet radiation, and the conditions for complete dechlorination were developed. Dechlorination was determined by analysis of the inorganic chloride formed and the concentrations of remaining PCBs. Transformer oil containing approximately 95 microg g(-1) PCB (approximately 3.5 mmol L(-1) Cl) is completely dechlorinated by irradiation with 600 kGy after the addition of 10% triethylamine (TEA). Transformer oil containing >800,000 microg g(-1) PCB (17.7 mol L(-1) Cl) requires an additional solvent to prevent solidification. When this oil is diluted with 2-propanol (2-PrOH) and TEA (v/v/v, 1/79/20), complete dechlorination is achieved with a dose of 2500 kGy. Ultraviolet photolysis of the same oil/2-PrOH/TEA solutions led to 90% dechlorination after exposure for 120 h in our experimental setup. Such yields were obtained by radiolysis with a dose of 2000 kGy (300 h in our Gammacell). Replacing TEA with KOH in 2-PrOH solutions greatly increases the yield of dechlorination in both the radiolytic and the photolytic experiments, demonstrating that a chain reaction plays a role in both of these treatment methods and suggesting that both methods deserve further consideration for large-scale application.

  14. Analytical methods for phenyltin compounds in polychlorinated biphenyl-based transformer oil samples.

    PubMed

    Yamamoto, Takashi; Noma, Yukio; Yasuhara, Akio; Sakai, Shin-ichi

    2003-10-31

    We present the first study on the analytical methods of phenyltin compounds (PTs) in polychlorinated biphenyl (PCB)-based transformer oil samples. Tetraphenyltin (TePhT) has been used as stabilizer for some kinds of PCBs-based transformer oil formulations. Monophenyltin (MPhT), diphenyltin (DPhT) and triphenyltin (TrPhT) could have been formed from TePhT during long-term use. TePhT was directly measured by gas chromatograph (GC) connected with three types of detectors, a mass spectrometer (MS), a flame photometric detector (FPD) and an atomic emission detector (AED) after dilution with hexane. MPhT, DPhT and TrPhT were propylated with Grignard reagent before measurement. The MS was the most sensitive of the detectors, with detection limits of phenyltin compounds of 30 ng/ml (MPhT), 9.8 ng/ml (DPhT), 5.5 ng/ml (TrPhT) and 0.60 ng/ml (TePhT), respectively. From the viewpoint of selectivity, MS was slightly worse than other detectors, but interference from PCBs matrices was not significant under ordinary analytical conditions. Two used transformer oil samples were analyzed using the analytical methods developed in this study. TePhT and TrPhT were found in both samples.

  15. Motion artifact reduction using hybrid Fourier transform with phase-shifting methods

    NASA Astrophysics Data System (ADS)

    Li, Beiwen; Liu, Ziping; Zhang, Song

    2016-08-01

    We propose to combine the Fourier transform profilometry (FTP) and phase-shifting profilometry (PSP) to reduce motion induced artifacts. The proposed method can be divided into three steps: Step 1 is to obtain a temporarily unwrapped absolute phase map of the entire scene using the FTP method, albeit the absolute phase map has motion introduced artifacts; Step 2 is to generate continuous relative phase maps without motion artifacts for each isolated object by spatially unwrapping each isolated phase map retrieved from the FTP method; and Step 3 is to determine the absolute phase map for each isolate region by referring to the temporally unwrapped phase using PSP method. Experimental results demonstrated success of the proposed method for measuring rapidly moving multiple isolated objects.

  16. Lifting of the Vlasov-Maxwell bracket by Lie-transform method

    NASA Astrophysics Data System (ADS)

    Brizard, A. J.; Morrison, P. J.; Burby, J. W.; de Guillebon, L.; Vittot, M.

    2016-12-01

    The Vlasov-Maxwell equations possess a Hamiltonian structure expressed in terms of a Hamiltonian functional and a functional bracket. In the present paper, the transformation (`lift') of the Vlasov-Maxwell bracket induced by the dynamical reduction of single-particle dynamics is investigated when the reduction is carried out by Lie-transform perturbation methods. The ultimate goal of this work is to provide an explicit pathway to the Hamiltonian formulations for the guiding-centre and gyrokinetic Vlasov-Maxwell equations, which have found important applications in our understanding of turbulent magnetized plasmas. Here, it is shown that the general form of the reduced Vlasov-Maxwell equations possesses a Hamiltonian structure defined in terms of a reduced Hamiltonian functional and a reduced bracket that automatically satisfies the standard bracket properties.

  17. [A phase error correction method for the new Fourier transforms spectrometer].

    PubMed

    Wang, Ning; Gong, Tian-Cheng; Chen, Jian-Jun; Li, Yang; Yang, Yi-Ning; Zhu, Yong; Zhang, Jie; Chen, Wei-Min

    2014-11-01

    To decrease the distortion of the recovered spectrum, improve the quantity of the recovered spectrum and decrease the influence of the phase error of the new spectrum detection system based on MEMS (micro-electro-mechanical systems) micro-mirrors, a new phase error correction method for this system is proposed in the present paper. The source of phase error of the spectrum detection system based on MEMS micro-mirrors is analyzed firstly. The analyzed result indicated that the phase error of the new spectral Fourier transform detection system is the zero drift of the optical path difference, and the phase error can be corrected by Zero-crossing sampling which is realized by improving the structure of the interferometer system and Mertz product The spectrum detection system is set up and the phase error correction method is verified by this system. The experiment result is show that the quantity of the recovered spectrum of the spectrum detection is improved obviously by using the improved interferometer system and Mertz product, and the recovered spectrum has no negative peaks and the side lobes is suppressed markedly. This correction method can reduce the influence caused by phase error to the system performance well and improve the spectral detection performance effectively. In this paper, the origin of the system phase error based on the new MEMS micromirror Fourier transform spectroscopy detection system is analyzed, and the phase error correction method is proposed. This method can improve the performance of the spectrum detection system.

  18. Cluster Variation Method as a Theoretical Tool for the Study of Phase Transformation

    NASA Astrophysics Data System (ADS)

    Mohri, Tetsuo

    2017-02-01

    Cluster variation method (CVM) has been widely employed to calculate alloy phase diagrams. The atomistic feature of the CVM is consistent with first-principles electronic structure calculations, and the combination of CVM with electronic structure calculation enables one to formulate free energy from the first-principles. CVM free energy conveys affluent information of a given system, and the second-order derivative traces the stability locus against configurational fluctuation. The kinetic extension of the CVM is the path probability method (PPM) which is utilized to calculate transformation and relaxation kinetics associated with the temperature change. Hence, the CVM and PPM are coherent methods to perform a synthetic study from initial non-equilibrium to final equilibrium states. By utilizing CVM free energy as a homogeneous free energy density term, one can calculate the time evolution of ordered domains within the phase field method. Finally, continuous displacement cluster variation method (CDCVM) is discussed as the recent development of CVM. CDCVM is capable of introducing the local lattice displacement into the free energy. Moreover, it is shown that CDCVM can be extended to study collective atomic displacements leading to displacive phase transformation.

  19. Improved Sensitivity Testing of Explosives Using Transformed Up-Down Methods

    NASA Astrophysics Data System (ADS)

    Brown, Geoffrey

    2013-06-01

    Sensitivity tests provide data that help establish guidelines for the safe handling of explosives. Any sensitivity test is based on assumptions to simplify the method or reduce the number of individual sample evaluations. Two common assumptions that are not typically checked after testing are 1) explosive response is linear in the applied stimulus levels and 2) the chosen test level spacing is close to the standard deviation of the explosive response function (for Bruceton Up-Down testing). In this work we present efforts to improve sensitivity testing by addressing these assumptions using Transformed Up-Down (TUD) test methods augmented with simple algorithms. TUD methods have been developed extensively for psychometric testing applications over the past 50 years and generally use multiple tests at a given level to determine whether to increase or decrease the applied stimulus. In the context of our work, we can use TUD methods that concentrate testing around useful probability levels and augment them with algorithms for adjusting test level spacing during the evaluation. Transformed methods that we have developed to address the assumptions noted above will be examined and evaluated with simulation to highlight their properties and assess their usefulness.

  20. Comprehensive reliability allocation method for CNC lathes based on cubic transformed functions of failure mode and effects analysis

    NASA Astrophysics Data System (ADS)

    Yang, Zhou; Zhu, Yunpeng; Ren, Hongrui; Zhang, Yimin

    2015-03-01

    Reliability allocation of computerized numerical controlled(CNC) lathes is very important in industry. Traditional allocation methods only focus on high-failure rate components rather than moderate failure rate components, which is not applicable in some conditions. Aiming at solving the problem of CNC lathes reliability allocating, a comprehensive reliability allocation method based on cubic transformed functions of failure modes and effects analysis(FMEA) is presented. Firstly, conventional reliability allocation methods are introduced. Then the limitations of direct combination of comprehensive allocation method with the exponential transformed FMEA method are investigated. Subsequently, a cubic transformed function is established in order to overcome these limitations. Properties of the new transformed functions are discussed by considering the failure severity and the failure occurrence. Designers can choose appropriate transform amplitudes according to their requirements. Finally, a CNC lathe and a spindle system are used as an example to verify the new allocation method. Seven criteria are considered to compare the results of the new method with traditional methods. The allocation results indicate that the new method is more flexible than traditional methods. By employing the new cubic transformed function, the method covers a wider range of problems in CNC reliability allocation without losing the advantages of traditional methods.

  1. Transformation of Lactuca sativa L. with rol C gene results in increased antioxidant potential and enhanced analgesic, anti-inflammatory and antidepressant activities in vivo.

    PubMed

    Ismail, Hammad; Dilshad, Erum; Waheed, Mohammad Tahir; Sajid, Moniba; Kayani, Waqas Khan; Mirza, Bushra

    2016-12-01

    Lettuce is an important edible crop which possesses various medicinal properties. In this study Lactuca sativa L. (cv Grand Rapids) was transformed by Agrobacterium-mediated transformation with rol C gene. Transgene integration and expression was confirmed through PCR and semiquantitative RT-PCR. The transformed extracts were evaluated for their in vitro antioxidant and in vivo analgesic, anti-inflammatory and antidepressant activities in rats. The transformed plants showed 53-98 % increase in total phenolic and 45-58 % increase in total flavonoid contents compared with untransformed plants. Results of total reducing power and total antioxidant capacity exhibited 90-118 and 61-75 % increase in transformed plants, respectively. In contrast to control, DPPH, lipid peroxidation and DNA protection assay showed up to 37, 20 and 50 % enhancement in transformed plants, respectively. The extracts showed similar but significant enhancement behavior in hot plate analgesic and carrageenan-induced hind paw edema test. The transformed extracts showed 72.1 and 78.5 % increase for analgesic and anti-inflammatory activities, respectively. The transformants of rol C gene exhibited prominent antidepressant activity with 64-73 % increase compared with untransformed plants. In conclusion, the present work suggests that transformation with rol C gene can be used to generate lettuce with enhanced medicinally important properties, such as antioxidant, analgesic, anti-inflammatory and antidepressant potential.

  2. Method of range profile for step frequency MMW radar based on wavelet transform power spectrum estimator

    NASA Astrophysics Data System (ADS)

    Li, Yuehua; Gao, Duntang; Shen, Qinghong; Li, Xingguo

    2001-11-01

    The method of range profile for step frequency MMW radar targets based on wavelet transform power spectrum estimator is studied. We show how the Fourier power spectrum can be detected by using the wavelet function coefficients (WFC) of the DWT. This method can successfully measure the power spectrum in samples for which traditional methods often fail because the sample are finite sized, have a complex geometry, or are varyingly sampled. We demonstrate that the spectrum features, such as the power law index, the magnitude, and the typical scales can be determined by the DWT reconstructed spectrum. We apply this method to the practical step frequency MMW radar target echo signals, and on the condition of the same sampling frequency and sampling data length, it can achieve one dimensional range profile with profile"s resolution superior to FFT"s, so the one dimensional range profile of targets can be analyzed with high resolution, the detail algorithm of range profiles spectrum estimation based on wavelet transforming multirange cells is proposed. Compare with FFT algorithm, using wavelet spectrum estimator of short data series, we can achieves high resolution, high accuracy, and low SNR threshold. The Experiment results make clear that the DWT estimator is a sensitive tool in range profile of step frequency MMW radar.

  3. Non-contact radio frequency shielding and wave guiding by multi-folded transformation optics method

    PubMed Central

    Madni, Hamza Ahmad; Zheng, Bin; Yang, Yihao; Wang, Huaping; Zhang, Xianmin; Yin, Wenyan; Li, Erping; Chen, Hongsheng

    2016-01-01

    Compared with conventional radio frequency (RF) shielding methods in which the conductive coating material encloses the circuits design and the leakage problem occurs due to the gap in such conductive material, non-contact RF shielding at a distance is very promising but still impossible to achieve so far. In this paper, a multi-folded transformation optics method is proposed to design a non-contact device for RF shielding. This “open-shielded” device can shield any object at a distance from the electromagnetic waves at the operating frequency, while the object is still physically open to the outer space. Based on this, an open-carpet cloak is proposed and the functionality of the open-carpet cloak is demonstrated. Furthermore, we investigate a scheme of non-contact wave guiding to remotely control the propagation of surface waves over any obstacles. The flexibilities of such multi-folded transformation optics method demonstrate the powerfulness of the method in the design of novel remote devices with impressive new functionalities. PMID:27841358

  4. [A novel method to determine the redshifts of active galaxies based on wavelet transform].

    PubMed

    Tu, Liang-Ping; Luo, A-Li; Jiang, Bin; Wei, Peng; Zhao, Yong-Heng; Liu, Rong

    2012-10-01

    Automatically determining redshifts of galaxies is very important for astronomical research on large samples, such as large-scale structure of cosmological significance. Galaxies are generally divided into normal galaxies and active galaxies, and the spectra of active galaxies mostly have more obvious emission lines. In the present paper, the authors present a novel method to determine spectral redshifts of active galaxies rapidly based on wavelet transformation mainly, and it does not need to extract line information accurately. This method includes the following steps: Firstly, we denoised a spectrum to be processed; Secondly, the low-frequency spectrum was extracted based on wavelet transform, and then we could get the residual spectrum through the denoised spectrum subtracting the low-frequency spectrum; Thirdly, the authors calculated the standard deviation of the residual spectrum and determined a threshold value T, then retained the wavelength set whose corresponding flux was greater than T; Fourthly, according to the wavelength form of all the standard lines, we calculated all the candidate redshifts; Finally, utilizing the density estimation method based on Parzen window, we determined the redshift point with maximum density, and the average value of its neighborhood would be the final redshift of this spectrum. The experiments on simulated data and real data from SDSS-DR7 show that this method is robust and its correct rate is encouraging. And it can be expected to be applied in the project of LAMOST.

  5. Non-singular acoustic cloak derived by the ray tracing method with rotationally symmetric transformations

    PubMed Central

    Wu, Linzhi

    2016-01-01

    Recently, the ray tracing method has been used to derive the non-singular cylindrical invisibility cloaks for out-of-plane shear waves, which is impossible via the transformation method directly owing to the singular push-forward mapping. In this paper, the method is adopted to design a kind of non-singular acoustic cloak. Based on Hamilton's equations of motion, eikonal equation and pre-designed ray equations, we derive several constraint equations for bulk modulus and density tensor. On the premise that the perfect matching conditions are satisfied, a series of non-singular physical profiles can be obtained by arranging the singular terms reasonably. The physical profiles derived by the ray tracing method will degenerate to the transformation-based solutions when taking the transport equation into consideration. This illuminates the essence of the newly designed cloaks that they are actually the so-called eikonal cloaks that can accurately control the paths of energy flux but with small disturbance in energy distribution along the paths. The near-perfect invisible performance has been demonstrated by the numerical ray tracing results and the pressure distribution snapshots. Finally, a kind of reduced cloak is conceived, and the good invisible performance has been measured quantitatively by the normalized scattering width. PMID:27118884

  6. Non-contact radio frequency shielding and wave guiding by multi-folded transformation optics method.

    PubMed

    Madni, Hamza Ahmad; Zheng, Bin; Yang, Yihao; Wang, Huaping; Zhang, Xianmin; Yin, Wenyan; Li, Erping; Chen, Hongsheng

    2016-11-14

    Compared with conventional radio frequency (RF) shielding methods in which the conductive coating material encloses the circuits design and the leakage problem occurs due to the gap in such conductive material, non-contact RF shielding at a distance is very promising but still impossible to achieve so far. In this paper, a multi-folded transformation optics method is proposed to design a non-contact device for RF shielding. This "open-shielded" device can shield any object at a distance from the electromagnetic waves at the operating frequency, while the object is still physically open to the outer space. Based on this, an open-carpet cloak is proposed and the functionality of the open-carpet cloak is demonstrated. Furthermore, we investigate a scheme of non-contact wave guiding to remotely control the propagation of surface waves over any obstacles. The flexibilities of such multi-folded transformation optics method demonstrate the powerfulness of the method in the design of novel remote devices with impressive new functionalities.

  7. Non-singular acoustic cloak derived by the ray tracing method with rotationally symmetric transformations.

    PubMed

    Gao, Penglin; Wu, Linzhi

    2016-02-01

    Recently, the ray tracing method has been used to derive the non-singular cylindrical invisibility cloaks for out-of-plane shear waves, which is impossible via the transformation method directly owing to the singular push-forward mapping. In this paper, the method is adopted to design a kind of non-singular acoustic cloak. Based on Hamilton's equations of motion, eikonal equation and pre-designed ray equations, we derive several constraint equations for bulk modulus and density tensor. On the premise that the perfect matching conditions are satisfied, a series of non-singular physical profiles can be obtained by arranging the singular terms reasonably. The physical profiles derived by the ray tracing method will degenerate to the transformation-based solutions when taking the transport equation into consideration. This illuminates the essence of the newly designed cloaks that they are actually the so-called eikonal cloaks that can accurately control the paths of energy flux but with small disturbance in energy distribution along the paths. The near-perfect invisible performance has been demonstrated by the numerical ray tracing results and the pressure distribution snapshots. Finally, a kind of reduced cloak is conceived, and the good invisible performance has been measured quantitatively by the normalized scattering width.

  8. A new modification to homotopy perturbation method combined with Fourier transform for solving nonlinear Cauchy reaction diffusion equation

    NASA Astrophysics Data System (ADS)

    Nourazar, S. S.; Nazari-Golshan, A.

    2015-01-01

    A hybrid of Fourier transform and new modified homotopy perturbation method based on the Adomian method is developed to solve linear and nonlinear partial differential equations. The Taylor series expansion is used to expand nonlinear term of partial differential equation and the Adomian polynomial incorporated into homotopy perturbation method combined with Fourier transform, is used to solve partial differential equations. Three case study problems, partial differential equations, are handled using homotopy perturbation method and Fourier transform modified homotopy perturbation method (FTMHPM). Results obtained are compared with exact solution. The comparison reveals that for same components of recursive sequences, errors associated with Fourier transform modified method are much less than the other and are valid for a large range of x-axis coordinates.

  9. Nanoscale displacement measurement by a digital nano-moiré method with wavelet transformation

    NASA Astrophysics Data System (ADS)

    Liu, Chia-Ming; Chen, Lien-Wen; Wang, Ching-Cheng

    2006-09-01

    A digital nano-moiré method with wavelet transformation is explored to measure nanoscale in-plane displacement fields. By applying e-beam lithography, a periodic PMMA nanostructure array is fabricated directly on the specimen and used as the specimen grating. Moiré patterns are generated by overlapping the images of the PMMA specimen grating obtained from AFM scanning and the virtual reference grating produced by a digital image generating process. Then, the overlapped images are filtered by the 2D wavelet transformation (WT) to capture the target moiré patterns. Existing methods, by overlapping the monitor-generated scanning lines with the image of the specimen grating, cause a mismatch problem. Previously, the carrier moiré method was explored with the aim of curing the mismatch problem. Unfortunately, the carrier moiré method, in addition to suffering from increased complexity of mathematical calculations, is incapable of directly obtaining the displacement field. Thus, the mismatch problem will result in inconveniences and restrictions in the practical application. Instead of using monitor-generated scanning lines, the proposed method applies the virtual reference grating, and thus puts the mismatch problem to rest. Nevertheless, the resultant moiré image suffers from low contrast which, if left untreated, might distort the measurement result. Therefore, the WT, known for its sharpened abilities of characteristic and edge detection, is used to capture the target moiré patterns and improve the measurement accuracy. The proposed method has been carried out in the laboratory. Experimental results have shown that the proposed method is convenient and efficient for nanoscale displacement measurement.

  10. Transform methods for precision continuum and control models of flexible space structures

    NASA Technical Reports Server (NTRS)

    Lupi, Victor D.; Turner, James D.; Chun, Hon M.

    1991-01-01

    An open loop optimal control algorithm is developed for general flexible structures, based on Laplace transform methods. A distributed parameter model of the structure is first presented, followed by a derivation of the optimal control algorithm. The control inputs are expressed in terms of their Fourier series expansions, so that a numerical solution can be easily obtained. The algorithm deals directly with the transcendental transfer functions from control inputs to outputs of interest, and structural deformation penalties, as well as penalties on control effort, are included in the formulation. The algorithm is applied to several structures of increasing complexity to show its generality.

  11. Method and apparatus for connecting high voltage leads to a high temperature super-conducting transformer

    DOEpatents

    Golner, Thomas M.; Mehta, Shirish P.

    2005-07-26

    A method and apparatus for connecting high voltage leads to a super-conducting transformer is provided that includes a first super-conducting coil set, a second super-conducting coil set, and a third super-conducting coil set. The first, second and third super-conducting coil sets are connected via an insulated interconnect system that includes insulated conductors and insulated connectors that are utilized to connect the first, second, and third super-conducting coil sets to the high voltage leads.

  12. An investigation of the kinetics of the mesophase transformations in petroleum pitch by the ESR method

    SciTech Connect

    Samoilov, V.M.; Frolov, V.I.; Kotosonov, A.S.; Ostronov, B.G.; Rumyantsev, S.M.

    1983-01-01

    The amount of ..cap alpha../sub 1/ fraction, the concentration of paramagnetic centers, and the width of the line of the ESR signal have been determined for samples of mesophase pitches obtained by treating an initial petroleum pitch at 400-440/sup 0/C. The applicability of the ESR method for studying the kinetics and mechanism of the process of mesophase transformations has been shown. From the change in the parameters studied the rate constants and activation energies of the processes of the formation and growth of the aromatic regions of polyconjugation and of the accumulation of mesophase have been calculated.

  13. Application of Distribution Transformer Thermal Life Models to Electrified Vehicle Charging Loads Using Monte-Carlo Method: Preprint

    SciTech Connect

    Kuss, M.; Markel, T.; Kramer, W.

    2011-01-01

    Concentrated purchasing patterns of plug-in vehicles may result in localized distribution transformer overload scenarios. Prolonged periods of transformer overloading causes service life decrements, and in worst-case scenarios, results in tripped thermal relays and residential service outages. This analysis will review distribution transformer load models developed in the IEC 60076 standard, and apply the model to a neighborhood with plug-in hybrids. Residential distribution transformers are sized such that night-time cooling provides thermal recovery from heavy load conditions during the daytime utility peak. It is expected that PHEVs will primarily be charged at night in a residential setting. If not managed properly, some distribution transformers could become overloaded, leading to a reduction in transformer life expectancy, thus increasing costs to utilities and consumers. A Monte-Carlo scheme simulated each day of the year, evaluating 100 load scenarios as it swept through the following variables: number of vehicle per transformer, transformer size, and charging rate. A general method for determining expected transformer aging rate will be developed, based on the energy needs of plug-in vehicles loading a residential transformer.

  14. A Fourier transform method for Vsin i estimations under nonlinear Limb-Darkening laws

    SciTech Connect

    Levenhagen, R. S.

    2014-12-10

    Star rotation offers us a large horizon for the study of many important physical issues pertaining to stellar evolution. Currently, four methods are widely used to infer rotation velocities, namely those related to line width calibrations, on the fitting of synthetic spectra, interferometry, and on Fourier transforms (FTs) of line profiles. Almost all of the estimations of stellar projected rotation velocities using the Fourier method in the literature have been addressed with the use of linear limb-darkening (LD) approximations during the evaluation of rotation profiles and their cosine FTs, which in certain cases, lead to discrepant velocity estimates. In this work, we introduce new mathematical expressions of rotation profiles and their Fourier cosine transforms assuming three nonlinear LD laws—quadratic, square-root, and logarithmic—and study their applications with and without gravity-darkening (GD) and geometrical flattening (GF) effects. Through an analysis of He I models in the visible range accounting for both limb and GD, we find out that, for classical models without rotationally driven effects, all the Vsin i values are too close to each other. On the other hand, taking into account GD and GF, the Vsin i obtained with the linear law result in Vsin i values that are systematically smaller than those obtained with the other laws. As a rule of thumb, we apply these expressions to the FT method to evaluate the projected rotation velocity of the emission B-type star Achernar (α Eri).

  15. A Fourier Transform Method for Vsin i Estimations under Nonlinear Limb-darkening Laws

    NASA Astrophysics Data System (ADS)

    Levenhagen, R. S.

    2014-12-01

    Star rotation offers us a large horizon for the study of many important physical issues pertaining to stellar evolution. Currently, four methods are widely used to infer rotation velocities, namely those related to line width calibrations, on the fitting of synthetic spectra, interferometry, and on Fourier transforms (FTs) of line profiles. Almost all of the estimations of stellar projected rotation velocities using the Fourier method in the literature have been addressed with the use of linear limb-darkening (LD) approximations during the evaluation of rotation profiles and their cosine FTs, which in certain cases, lead to discrepant velocity estimates. In this work, we introduce new mathematical expressions of rotation profiles and their Fourier cosine transforms assuming three nonlinear LD laws—quadratic, square-root, and logarithmic—and study their applications with and without gravity-darkening (GD) and geometrical flattening (GF) effects. Through an analysis of He I models in the visible range accounting for both limb and GD, we find out that, for classical models without rotationally driven effects, all the Vsin i values are too close to each other. On the other hand, taking into account GD and GF, the Vsin i obtained with the linear law result in Vsin i values that are systematically smaller than those obtained with the other laws. As a rule of thumb, we apply these expressions to the FT method to evaluate the projected rotation velocity of the emission B-type star Achernar (α Eri).

  16. Simulations of string vibrations with boundary conditions of third kind using the functional transformation method

    NASA Astrophysics Data System (ADS)

    Trautmann, L.; Petrausch, S.; Bauer, M.

    2005-09-01

    The functional transformation method (FTM) is an established mathematical method for accurate simulation of multidimensional physical systems from various fields of science, including optics, heat and mass transfer, electrical engineering, and acoustics. It is a frequency-domain method based on the decomposition into eigenvectors and eigenfrequencies of the underlying physical problem. In this article, the FTM is applied to real-time simulations of vibrating strings which are ideally fixed at one end while the fixing at the other end is modeled by a frequency-dependent input impedance. Thus, boundary conditions of third kind are applied to the model at the end fixed with the input impedance. It is shown that accurate and stable simulations are achieved with nearly the same computational cost as with strings ideally fixed at both ends.

  17. Multiscale characterization method for line edge roughness based on redundant second generation wavelet transform

    SciTech Connect

    Wang Fei; Zhao Xuezeng; Li Ning

    2010-10-15

    We introduce a multiscale characterization method for line edge roughness (LER) based on redundant second generation wavelet transform. This method involves decomposing LER characteristics into independent bands with different spatial frequency components at different scales, and analyzing the reconstructed signals to work out the roughness exponent, the spatial frequency distribution characteristics, as well as the rms value. The effect of noise can be predicted using detailed signals in the minimum space of scale. This method was applied to numerical profiles for validation. Results show that according to the line edge profiles with similar amplitudes, the roughness exponent R can effectively reflect the degree of irregularity of LER and intuitively provide information on LER spatial frequency distribution.

  18. General properties of the Foldy-Wouthuysen transformation and applicability of the corrected original Foldy-Wouthuysen method

    NASA Astrophysics Data System (ADS)

    Silenko, Alexander J.

    2016-02-01

    General properties of the Foldy-Wouthuysen transformation which is widely used in quantum mechanics and quantum chemistry are considered. Merits and demerits of the original Foldy-Wouthuysen transformation method are analyzed. While this method does not satisfy the Eriksen condition of the Foldy-Wouthuysen transformation, it can be corrected with the use of the Baker-Campbell-Hausdorff formula. We show a possibility of such a correction and propose an appropriate algorithm of calculations. An applicability of the corrected Foldy-Wouthuysen method is restricted by the condition of convergence of a series of relativistic corrections.

  19. Ovary-drip transformation: a simple method for directly generating vector- and marker-free transgenic maize (Zea mays L.) with a linear GFP cassette transformation.

    PubMed

    Yang, Aifu; Su, Qiao; An, Lijia

    2009-03-01

    The presence of selectable marker genes and vector backbone sequences has affected the safe assessment of transgenic plants. In this study, the ovary-drip method for directly generating vector- and selectable marker-free transgenic plants was described, by which maize was transformed with a linear GFP cassette (Ubi-GFP-nos). The key features of this method center on the complete removal of the styles and the subsequent application of a DNA solution directly to the ovaries. The movement of the exogenous DNA was monitored using fluorescein isothiocyanate-labeled DNA, which showed that the time taken by the exogenous DNA to enter the ovaries was shortened compared to that of the pollen-tube pathway. This led to an improved transformation frequency of 3.38% compared to 0.86% for the pollen-tube pathway as determined by PCR analysis. The use of 0.05% surfactant Silwet L-77 + 5% sucrose as a transformation solution further increased the transformation frequency to 6.47%. Southern blot analysis showed that the transgenic plants had low transgene copy number and simple integration pattern. Green fluorescence was observed in roots and immature embryos of transgenic plants by fluorescence microscopy. Progeny analysis showed that GFP insertions were inherited in T(1) generation. The ovary-drip method would become a favorable choice for directly generating vector- and marker-free transgenic maize expressing functional genes of agronomic interest.

  20. Improved sensitivity testing of explosives using transformed Up-Down methods

    NASA Astrophysics Data System (ADS)

    Brown, Geoffrey W.

    2014-05-01

    Sensitivity tests provide data that help establish guidelines for the safe handling of explosives. Any sensitivity test is based on assumptions to simplify the method or reduce the number of individual sample evaluations. Two common assumptions that are not typically checked after testing are 1) explosive response follows a normal distribution as a function of the applied stimulus levels and 2) the chosen test level spacing is close to the standard deviation of the explosive response function (for Bruceton Up-Down testing for example). These assumptions and other limitations of traditional explosive sensitivity testing can be addressed using Transformed Up-Down (TUD) test methods. TUD methods have been developed extensively for psychometric testing over the past 50 years and generally use multiple tests at a given level to determine how to adjust the applied stimulus. In the context of explosive sensitivity we can use TUD methods that concentrate testing around useful probability levels. Here, these methods are explained and compared to Bruceton Up-Down testing using computer simulation. The results show that the TUD methods are more useful for many cases but that they do require more tests as a consequence. For non-normal distributions, however, the TUD methods may be the only accurate assessment method.

  1. Development of an improved analytical method for the determination of carcinogenic polycyclic aromatic hydrocarbons in transformer oil.

    PubMed

    Pillai, I; Ritchie, L; Heywood, R; Wilson, G; Pahlavanpour, B; Setford, S; Saini, S

    2005-02-04

    Polynuclear aromatic hydrocarbons (PAHs) are natural constituents of transformer oils and are essential in prolonging transformer in-service lifetime. Issues concerning PAH carcinogenicity demand methods that provide qualitative and quantitative information on the PAH composition of new and in-service oils to allow informed operational decisions to be made. However, current analytical methods focus on PAH fingerprinting, as opposed to quantitative analysis and are also cumbersome, relying on the use of large (>100 ml) volumes of organic solvents, some of which are hazardous. This paper reports a method for the improved quantification of carcinogenic PAHs in transformer oils that is both simple and repeatable. The method uses commercially available solid-phase extraction columns and millilitre volumes of relatively non-hazardous solvents. Extraction efficiencies of > or =74% were obtained for the Environmental Protection Agency priority PAHs. The method has potential for automation and high-throughput analysis and thus is of interest to industries that use transformer oils.

  2. The Empirical Comparison of Coordinate Transformation Models and Distortion Modeling Methods Based on a Case Study of Croatia

    NASA Astrophysics Data System (ADS)

    Grgic, M.; Varga, M.; Bašić, T.

    2015-12-01

    Several coordinate transformation models enable performing of the coordinate transformations between the historical astro-geodetic datums, which were utilized before the GNSS (Global Navigation Satellite System) technologies were developed, and datums related to the International Terrestrial Reference System (ITRS), which today are most often used to determine the position. The decision on the most appropriate coordinate transformation model is influenced by many factors, such as: required accuracy, available computational resources, possibility of the model application regarding the size and shape of the territory, coordinate distortion that very often exist in historical astro-geodetic datums, etc. This study is based on the geodetic data of the Republic of Croatia in both, historical and ITRS-related datum. It investigates different transformation models, including conformal Molodensky 3 parameters (p) and 5p (standard and abridged) transformation models, 7p transformation models (Bursa-Wolf and Molodensky-Badekas model), Affine transformation models (8p, 9p, 12p), and Multiple Regression Equation approach. Besides, it investigates the 7p, 8p, 9p, and 12p transformation models extended with distortion modeling, and the grid based only transformation model (NTv2 model). Furthermore, several distortion modeling methods were used to produce various models of distortion shifts in different resolutions. Thereafter, their performance and the performance of the transformation models was evaluated using summary statistics derived from the remained positional residuals that were computed for the independent control spatial data set. Lastly, the most appropriate method(s) of distortion modeling and most appropriate coordinate transformation model(s) were defined regarding the required accuracy for the Croatian case.

  3. New method for the transformation of solar radiation energy into electric power for energy feeding of the space vehicles

    NASA Astrophysics Data System (ADS)

    Ludanov, K. I.

    The author proposes a new method for the transformation of solar radiation energy into electric power, which is alternative for photo-transformation. Ukrpatents's positive decisions are obtained for the method and for the installation for its realization. The method includes two phases: concentration of solar radiation by paraboloid mirrors with high potential heat obtaining in the helio receiver and the next heat transformation into electric power in the framework of the thermal cycle "high temperature electrolytic steam decomposition on the components (H2 and O2) + electrochemical generation by the way of the water recombination from H2 and O2 in the low temperature fuel cell". The new method gives the double superiority in comparison with the photo-transformation.

  4. A coordinate transformation method for calculating the 3D light intensity distribution in ICF hohlraum

    NASA Astrophysics Data System (ADS)

    Lin, Zhili; Li, Xiaoyan; Zhao, Kuixia; Chen, Xudong; Chen, Mingyu; Pu, Jixiong

    2016-06-01

    For an inertial confinement fusion (ICF) system, the light intensity distribution in the hohlraum is key to the initial plasma excitation and later laser-plasma interaction process. Based on the concept of coordinate transformation of spatial points and vector, we present a robust method with a detailed procedure that makes the calculation of the three dimensional (3D) light intensity distribution in hohlraum easily. The method is intuitive but powerful enough to solve the complex cases of random number of laser beams with arbitrary polarization states and incidence angles. Its application is exemplified in the Shenguang III Facility (SG-III) that verifies its effectiveness and it is useful for guiding the design of hohlraum structure parameter.

  5. Multirate Simulations of String Vibrations Including Nonlinear Fret-String Interactions Using the Functional Transformation Method

    NASA Astrophysics Data System (ADS)

    Trautmann, L.; Rabenstein, R.

    2004-12-01

    The functional transformation method (FTM) is a well-established mathematical method for accurate simulations of multidimensional physical systems from various fields of science, including optics, heat and mass transfer, electrical engineering, and acoustics. This paper applies the FTM to real-time simulations of transversal vibrating strings. First, a physical model of a transversal vibrating lossy and dispersive string is derived. Afterwards, this model is solved with the FTM for two cases: the ideally linearly vibrating string and the string interacting nonlinearly with the frets. It is shown that accurate and stable simulations can be achieved with the discretization of the continuous solution at audio rate. Both simulations can also be performed with a multirate approach with only minor degradations of the simulation accuracy but with preservation of stability. This saves almost 80% of the computational cost for the simulation of a six-string guitar and therefore it is in the range of the computational cost for digital waveguide simulations.

  6. Reference sample method reduces the error caused by variable cryosection thickness in Fourier transform infrared imaging.

    PubMed

    Rieppo, Jarno; Hyttinen, Mika M; Jurvelin, Jukka S; Helminen, Heikki J

    2004-01-01

    Fourier transform infrared imaging (FT-IRI) is a novel technique for characterization of the biochemical composition of biological tissues, e.g., articular cartilage. The use of cryosections is preferred in FT-IRI. Unfortunately, significant variation in section thickness often impairs the suitability of cryosections for quantitative FT-IRI analysis. The present study introduces an inexpensive reference sample method for quantitative analysis. In this technique, specimen absorption is normalized with that of nitrocellulose membrane embedded and cryosectioned with the sample. Mean variation of the infrared absorption in cartilage specimens was 11.5%, 12.1%, and 20.6% for 5 microm, 10 microm, and 14 microm thick sections, respectively, without normalization. Normalization reduced the variation to 5.2%, 4.0%, and 4.6% for the same sections, respectively. The normalization method enables usage of cryosections for quantitative work and significantly reduces the cost and time needed for FT-IRI analysis.

  7. A novel transform domain processing based channel estimation method for OFDM radio-over-fiber systems.

    PubMed

    Tao, Li; Yu, Jianjun; Yang, Qi; Shao, Yufeng; Zhang, Junwen; Chi, Nan

    2013-03-25

    In this paper, a transform domain processing (TDP) based channel estimation method for orthogonal frequency-division multiplexing (OFDM) Radio-over-Fiber (RoF) systems is proposed. Theoretically investigation shows that TDP can greatly reduce the number of required training symbols. An 8 x 4.65 Gb/s multi-user OFDM RoF system over 40 km fiber link and 60 GHz wireless link is experimentally demonstrated utilizing TDP scheme. Compared with conventional time domain averaging (TDA) scheme, the overhead can be reduced from several tens of training symbols to merely one symbol and the receiver sensitivity has been improved by 1.8 dB at BER of 3.8 x 10(-3). The calculated BER performance for 8 wireless users clearly validates the feasibility of this TDP-based channel estimation method.

  8. Third-order symplectic integration method with inverse time dispersion transform for long-term simulation

    NASA Astrophysics Data System (ADS)

    Gao, Yingjie; Zhang, Jinhai; Yao, Zhenxing

    2016-06-01

    The symplectic integration method is popular in high-accuracy numerical simulations when discretizing temporal derivatives; however, it still suffers from time-dispersion error when the temporal interval is coarse, especially for long-term simulations and large-scale models. We employ the inverse time dispersion transform (ITDT) to the third-order symplectic integration method to reduce the time-dispersion error. First, we adopt the pseudospectral algorithm for the spatial discretization and the third-order symplectic integration method for the temporal discretization. Then, we apply the ITDT to eliminate time-dispersion error from the synthetic data. As a post-processing method, the ITDT can be easily cascaded in traditional numerical simulations. We implement the ITDT in one typical exiting third-order symplectic scheme and compare its performances with the performances of the conventional second-order scheme and the rapid expansion method. Theoretical analyses and numerical experiments show that the ITDT can significantly reduce the time-dispersion error, especially for long travel times. The implementation of the ITDT requires some additional computations on correcting the time-dispersion error, but it allows us to use the maximum temporal interval under stability conditions; thus, its final computational efficiency would be higher than that of the traditional symplectic integration method for long-term simulations. With the aid of the ITDT, we can obtain much more accurate simulation results but with a lower computational cost.

  9. Regulation of astaxanthin and its intermediates through cloning and genetic transformation of β-carotene ketolase in Haematococcus pluvialis.

    PubMed

    Kathiresan, S; Chandrashekar, Arun; Ravishankar, G A; Sarada, R

    2015-02-20

    Astaxanthin, a high-value ketocarotenoid used in the pharmaceutical and nutraceutical industries is mainly produced from green alga, Haematococcus pluvialis. It is biosynthesized by the action of key enzyme, β-carotene ketolase (BKT) on β-carotene through intermediates echinenone and canthaxanthin. In this study, the β-carotene ketolase (bkt) gene was isolated from H. pluvialis and cloned in a vector pRT100 and further mobilized to a binary vector pCAMBIA 1304. The T-DNA of pCAMBIA 1304, which consists of cloned bkt, was successfully transformed to H. pluvialis through Agrobacterium mediation. The cloning and transformation of bkt in H. pluvialis was confirmed by Southern blotting and also by PCR analysis. Total carotenoids and astaxanthin content in the transformed cells were found to be 2-3-fold higher, while the intermediates like echinenone and canthaxanthin were found to be 8-10-fold higher than in the control cells. The expression level of carotenogenic genes like phytoene synthase (psy), phytoene desaturase (pds), lycopene cyclase (lcy), bkt, and β-carotene hydroxylase (bkh) were found to be higher in transformed cells compared to the non-transformed (NT) H. pluvialis.

  10. Development of an efficient regeneration and transformation method for the new potential oilseed crop Lepidium campestre

    PubMed Central

    2013-01-01

    Background Lepidium campestre is an undomesticated oilseed species with a great potential to become a new crop for both food and industrial feedstocks production. Genetic modification is needed for further improving the oil quantity and quality of Lepidium. Studies on in vitro shoot regeneration of Lepidium are very limited and there is no transformation protocol available. Results We have investigated the effects of different factors, especially the type, concentration and combination of plant growth regulators (PGRs) on in vitro shoot regeneration of Lepidium. The results showed that the 2,4-D treatment was crucial to shoot regeneration from different explants. The duration of 2,4-D exposure between 2-4 days did not show significant difference in shoot regeneration, while the effect of 2,4-D concentration varied greatly depending on the type of explants and cytokinins used, for example, the low concentration of 2,4-D combined with TDZ significantly increased the regeneration frequency of hypocotyls. Cotyledon and hypocotyl explants responded differently to cytokinin, for example, TDZ was more effective than zeatin in promoting shoot regeneration from hypocotyls, but did not affect the regeneration of cotyledons which was more affected by high concentration of zeatin. The results also showed that NAA was not effective for shoot regeneration. Germination in light increased the regeneration frequency compared to that in dark. After optimization of the different conditions, an efficient regeneration protocol was developed with the regeneration efficiency of 92.7%. Using this protocol, the transformation frequency of 6% in average was achieved. The presence of transgenes in the transgenic lines was confirmed by GUS staining, PCR and Southern blot analyses. Conclusion Through systematic investigation of important factors affecting in vitro shoot regeneration, we have developed an efficient regeneration and transformation protocol for the genetic modification of

  11. Comparison of automatic denoising methods for phonocardiograms with extraction of signal parameters via the Hilbert Transform

    NASA Astrophysics Data System (ADS)

    Messer, Sheila R.; Agzarian, John; Abbott, Derek

    2001-05-01

    Phonocardiograms (PCGs) have many advantages over traditional auscultation (listening to the heart) because they may be replayed, may be analyzed for spectral and frequency content, and frequencies inaudible to the human ear may be recorded. However, various sources of noise may pollute a PCG including lung sounds, environmental noise and noise generated from contact between the recording device and the skin. Because PCG signals are known to be nonlinear and it is often not possible to determine their noise content, traditional de-noising methods may not be effectively applied. However, other methods including wavelet de-noising, wavelet packet de-noising and averaging can be employed to de-noise the PCG. This study examines and compares these de-noising methods. This study answers such questions as to which de-noising method gives a better SNR, the magnitude of signal information that is lost as a result of the de-noising process, the appropriate uses of the different methods down to such specifics as to which wavelets and decomposition levels give best results in wavelet and wavelet packet de-noising. In general, the wavelet and wavelet packet de-noising performed roughly equally with optimal de-noising occurring at 3-5 levels of decomposition. Averaging also proved a highly useful de- noising technique; however, in some cases averaging is not appropriate. The Hilbert Transform is used to illustrate the results of the de-noising process and to extract instantaneous features including instantaneous amplitude, frequency, and phase.

  12. Verification and Validation of a Coordinate Transformation Method in Axisymmetric Transient Magnetics.

    SciTech Connect

    Ashcraft, C. Chace; Niederhaus, John Henry; Robinson, Allen C.

    2016-01-29

    We present a verification and validation analysis of a coordinate-transformation-based numerical solution method for the two-dimensional axisymmetric magnetic diffusion equation, implemented in the finite-element simulation code ALEGRA. The transformation, suggested by Melissen and Simkin, yields an equation set perfectly suited for linear finite elements and for problems with large jumps in material conductivity near the axis. The verification analysis examines transient magnetic diffusion in a rod or wire in a very low conductivity background by first deriving an approximate analytic solution using perturbation theory. This approach for generating a reference solution is shown to be not fully satisfactory. A specialized approach for manufacturing an exact solution is then used to demonstrate second-order convergence under spatial refinement and tem- poral refinement. For this new implementation, a significant improvement relative to previously available formulations is observed. Benefits in accuracy for computed current density and Joule heating are also demonstrated. The validation analysis examines the circuit-driven explosion of a copper wire using resistive magnetohydrodynamics modeling, in comparison to experimental tests. The new implementation matches the accuracy of the existing formulation, with both formulations capturing the experimental burst time and action to within approximately 2%.

  13. Lifting of the Vlasov-Maxwell Bracket by Lie-transform Method

    NASA Astrophysics Data System (ADS)

    Brizard, A. J.; Morrison, P. J.; Burby, J. W.

    2016-10-01

    The Vlasov-Maxwell equations possess a Hamiltonian structure expressed in terms of a Hamiltonian functional and a functional bracket. The transformation (``lift'') of the Vlasov-Maxwell bracket induced by the dynamical reduction of single-particle dynamics is investigated when the reduction is carried out by Lie-transform perturbation methods. The ultimate goal of this work is to derive explicit Hamiltonian formulations for the guiding-center and gyrokinetic Vlasov-Maxwell equations that have important applications in our understanding of turbulent magnetized plasmas. In particular, we investigate how the Hamiltonian properties of the reduced Vlasov-Maxwell bracket survive (1) the closure problem: the process of truncation of the guiding-center Vlasov-Maxwell bracket at a finite order in ɛ (so far expressions have been derived at all orders in ɛ) and (2) the averaging problem: the process by which which the gyroangle is eliminated from the guiding-center Vlasov-Maxwell bracket (since guiding-center Vlasov-Maxwell equations do not involve the fast gyromotion time scale). Work supported by Grants from US DoE.

  14. High-throughput transformation method for Yarrowia lipolytica mutant library screening.

    PubMed

    Leplat, Christophe; Nicaud, Jean-Marc; Rossignol, Tristan

    2015-09-01

    As a microorganism of major biotechnological importance, the oleaginous yeast Yarrowia lipolytica is subjected to intensive genetic engineering and functional genomic analysis. Future advancements in this area, however, require a system that will generate a large collection of mutants for high-throughput screening. Here, we report a rapid and efficient method for high-throughput transformation of Y. lipolytica in 96-well plates. We developed plasmids and strains for the large-scale screening of overexpression mutant strains, using Gateway® vectors that were adapted for specific locus integration in Y. lipolytica. As an example, a collection of mutants that overexpressed the alkaline extracellular protease (AEP) was obtained in a single transformation experiment. The platform strain that we developed to receive the overexpression cassette was designed to constitutively express a fluorescent protein as a convenient growth reporter for screening in non-translucid media. An example of growth comparison in skim milk-based medium between AEP overexpression and deletion mutants is provided.

  15. Method for simulating dose reduction in digital mammography using the Anscombe transformation

    PubMed Central

    Borges, Lucas R.; de Oliveira, Helder C. R.; Nunes, Polyana F.; Bakic, Predrag R.; Maidment, Andrew D. A.; Vieira, Marcelo A. C.

    2016-01-01

    Purpose: This work proposes an accurate method for simulating dose reduction in digital mammography starting from a clinical image acquired with a standard dose. Methods: The method developed in this work consists of scaling a mammogram acquired at the standard radiation dose and adding signal-dependent noise. The algorithm accounts for specific issues relevant in digital mammography images, such as anisotropic noise, spatial variations in pixel gain, and the effect of dose reduction on the detective quantum efficiency. The scaling process takes into account the linearity of the system and the offset of the detector elements. The inserted noise is obtained by acquiring images of a flat-field phantom at the standard radiation dose and at the simulated dose. Using the Anscombe transformation, a relationship is created between the calculated noise mask and the scaled image, resulting in a clinical mammogram with the same noise and gray level characteristics as an image acquired at the lower-radiation dose. Results: The performance of the proposed algorithm was validated using real images acquired with an anthropomorphic breast phantom at four different doses, with five exposures for each dose and 256 nonoverlapping ROIs extracted from each image and with uniform images. The authors simulated lower-dose images and compared these with the real images. The authors evaluated the similarity between the normalized noise power spectrum (NNPS) and power spectrum (PS) of simulated images and real images acquired with the same dose. The maximum relative error was less than 2.5% for every ROI. The added noise was also evaluated by measuring the local variance in the real and simulated images. The relative average error for the local variance was smaller than 1%. Conclusions: A new method is proposed for simulating dose reduction in clinical mammograms. In this method, the dependency between image noise and image signal is addressed using a novel application of the Anscombe

  16. Spectral transform methods for solving the shallow-water equations on the sphere

    SciTech Connect

    Swarztrauber, P.N.

    1996-04-01

    The accuracy of computed solutions to several formulations of the shallow-water equations is compared. The shallow-water equations can be written in a number of different forms that are obtained by (a) combining terms into differential expressions; (b) raising the order of the differential equations, for example, the vorticity-divergence formulation, and (c) transforming both independent and dependent variables. Although the exact solutions to the formulations are identical, the properties of the computed solutions vary depending on the formulation and the method of solution. Nine methods are examined, all of which provide satisfactory accuracy for a steady-state test case. The exact solution corresponds to a steady zonal wind that is tilted relative to the computational spherical coordinate system. The resulting wind passes over the North Pole, providing a test of the `pole problem.` Computational details are presented as well as the accuracy of a 5-day computation in both 32- and 64-bit arithmetic. Stability and accuracy are shown to depend on the polar smoothing induced by resynthesis, which is also the accepted method for eliminating nonlinear aliasing. Eight of the nine methods mimic the exact solution by computing a numerical steady-state solution. 19 refs., 2 figs., 4 tabs.

  17. A novel analysis method for near infrared spectroscopy based on Hilbert-Huang transform

    NASA Astrophysics Data System (ADS)

    Zhou, Zhenyu; Yang, Hongyu; Liu, Yun; Ruan, Zongcai; Luo, Qingming; Gong, Hui; Lu, Zuhong

    2007-05-01

    Near Infrared Imager (NIRI) has been widely used to access the brain functional activity non-invasively. We use a portable, multi-channel and continuous-wave NIR topography instrument to measure the concentration changes of each hemoglobin species and map cerebral cortex functional activation. By extracting some essential features from the BOLD signals, optical tomography is able to be a new way of neuropsychological studies. Fourier spectral analysis provides a common framework for examining the distribution of global energy in the frequency domain. However, this method assumes that the signal should be stationary, which limits its application in non-stationary system. The hemoglobin species concentration changes are of such kind. In this work we develop a new signal processing method using Hilbert-Huang transform to perform spectral analysis of the functional NIRI signals. Compared with wavelet based multi-resolution analysis (MRA), we demonstrated the extraction of task related signal for observation of activation in the prefrontal cortex (PFC) in vision stimulation experiment. This method provides a new analysis tool for functional NIRI signals. Our experimental results show that the proposed approach provides the unique method for reconstructing target signal without losing original information and enables us to understand the episode of functional NIRI more precisely.

  18. A new method to calculate the beam charge for an integrating current transformer

    SciTech Connect

    Wu Yuchi; Han Dan; Zhu Bin; Dong Kegong; Tan Fang; Gu Yuqiu

    2012-09-15

    The integrating current transformer (ICT) is a magnetic sensor widely used to precisely measure the charge of an ultra-short-pulse charged particle beam generated by traditional accelerators and new laser-plasma particle accelerators. In this paper, we present a new method to calculate the beam charge in an ICT based on circuit analysis. The output transfer function shows an invariable signal profile for an ultra-short electron bunch, so the function can be used to evaluate the signal quality and calculate the beam charge through signal fitting. We obtain a set of parameters in the output function from a standard signal generated by an ultra-short electron bunch (about 1 ps in duration) at a radio frequency linear electron accelerator at Tsinghua University. These parameters can be used to obtain the beam charge by signal fitting with excellent accuracy.

  19. A Novel Generation Method of Dielectric Barrier Discharge and Ozone Production Using a Piezoelectric Transformer

    NASA Astrophysics Data System (ADS)

    Teranishi, Kenji; Suzuki, Susumu; Itoh, Haruo

    2004-09-01

    A novel generation method of a dielectric barrier discharge (DBD) using a piezoelectric transformer (PT) is proposed. Spatio-temporal variations of microdischarges were investigated and discussed on the basis of the observations using a charge-coupled device (CCD) camera. As an example of a practical application of the DBD to a plasma reactor, an ozonizer using the PT was developed and ozone productions were performed in air and oxygen. Maximum ozone concentrations of 3.38 and 20.3 g/Nm3 were obtained in air and oxygen, respectively. The ozonizer can be designed in a compact configuration and driven with low applied voltages because the PT serves as both a high-voltage generator and a discharge electrode.

  20. A hybrid group method of data handling with discrete wavelet transform for GDP forecasting

    NASA Astrophysics Data System (ADS)

    Isa, Nadira Mohamed; Shabri, Ani

    2013-09-01

    This study is proposed the application of hybridization model using Group Method of Data Handling (GMDH) and Discrete Wavelet Transform (DWT) in time series forecasting. The objective of this paper is to examine the flexibility of the hybridization GMDH in time series forecasting by using Gross Domestic Product (GDP). A time series data set is used in this study to demonstrate the effectiveness of the forecasting model. This data are utilized to forecast through an application aimed to handle real life time series. This experiment compares the performances of a hybrid model and a single model of Wavelet-Linear Regression (WR), Artificial Neural Network (ANN), and conventional GMDH. It is shown that the proposed model can provide a promising alternative technique in GDP forecasting.

  1. Isospin-violating nucleon-nucleon forces using the method of unitary transformation

    SciTech Connect

    Evgeny Epelbaum; Ulf-G. Meissner

    2005-02-01

    Recently, we have derived the leading and subleading isospin-breaking three-nucleon forces using the method of unitary transformation. In the present work we extend this analysis and consider the corresponding two-nucleon forces using the same approach. Certain contributions to the isospin-violating one- and two-pion exchange potential have already been discussed by various groups within the effective field theory framework. Our findings agree with the previously obtained results. In addition, we present the expressions for the subleading charge-symmetry-breaking two-pion exchange potential which were not considered before. These corrections turn out to be numerically important. Together with the three-nucleon force results presented in our previous work, the results of the present study specify completely isospin-violating nuclear force up to the order {Lambda}{sup 5}.

  2. A quantitative and qualitative method to control chemotherapeutic preparations by Fourier transform infrared-ultraviolet spectrophotometry.

    PubMed

    Dziopa, Florian; Galy, Guillaume; Bauler, Stephanie; Vincent, Benoit; Crochon, Sarah; Tall, Mamadou Lamine; Pirot, Fabrice; Pivot, Christine

    2013-06-01

    Chemotherapy products in hospitals include a reconstitution step of manufactured drugs providing an adapted dosage to each patient. The administration of highly iatrogenic drugs raises the question of patients' safety and treatment efficiency. In order to reduce administration errors due to faulty preparations, we introduced a new qualitative and quantitative routine control based on Fourier Transform Infrared (FTIR) and UV-Visible spectrophotometry. This automated method enabled fast and specific control for 14 anticancer drugs. A 1.2 mL sample was used to assay and identify each preparation in less than 90 sec. Over a two-year period, 9370 controlled infusion bags showed a 1.49% nonconformity rate, under 15% tolerance from the theoretical concentration and 96% minimum identification matching factor. This study evaluated the reliability of the control process, as well as its accordance to chemotherapy deliverance requirements. Thus, corrective measures were defined to improve the control process.

  3. Application of the contact transformation method to torsional problems in methyl silane

    NASA Astrophysics Data System (ADS)

    Moazzen-Ahmadi, N.; Ozier, I.

    1987-11-01

    The effect of the torsional degree of freedom on redundancies in the Hamiltonian and on the dipole operator has been investigated for methyl silane-like molecules. By applying a rotational contact transformation to the torsion-rotation Hamiltonian HTR for the ground vibrational state, a systematic method is demonstrated for treating the redundancies that relate different terms in HTR. In general, with this method, the experimentally accessible molecular parameters in the reduced Hamiltonian can be related to the physically significant molecular parameters in the untransformed Hamiltonian. It is shown that HTR contains a new term which has matrix elements with selection rules (Δ K = ±3), (Δ σ = 0), and Δ vT arbitrary, where vT and σ label the torsional levels and sublevels, respectively. As a result of this term, the distortion dipole constant μD which characterizes (Δ K = ±3) matrix elements in C3 v molecules cannot, in systems like CH 3SiH 3, be ascribed entirely to centrifugal distortion but can contain a significant contribution from torsional effects. Furthermore, new transitions can appear in the pure torsional bands which may be strong enough to observe in low barrier molecules. By applying a vibrational contact transformation, the form is derived of the leading torsional terms in the dipole moment expansion. The four dipole distortion constants μ0T, μ2T, μ|;T, and μΛT which characterize these terms are related to the molecular parameters that enter the Coriolis, centrifugal distortion, and anharmonicity contributions to the vibration-torsion-rotation Hamiltonian.

  4. A combined compensation method for the output voltage of an insulated core transformer power supply

    SciTech Connect

    Yang, L.; Yang, J. Liu, K. F.; Qin, B.; Chen, D. Z.

    2014-06-15

    An insulated core transformer (ICT) power supply is an ideal high-voltage generator for irradiation accelerators with energy lower than 3 MeV. However, there is a significant problem that the structure of the segmented cores leads to an increase in the leakage flux and voltage differences between rectifier disks. A high level of consistency in the output of the disks helps to achieve a compact structure by improving the utilization of both the rectifier components and the insulation distances, and consequently increase the output voltage of the power supply. The output voltages of the disks which are far away from the primary coils need to be improved to reduce their inhomogeneity. In this study, by investigating and comparing the existing compensation methods, a new combined compensation method is proposed, which increases the turns on the secondary coils and employs parallel capacitors to improve the consistency of the disks, while covering the entire operating range of the power supply. This method turns out to be both feasible and effective during the development of an ICT power supply. The non-uniformity of the output voltages of the disks is less than 3.5% from no-load to full-load, and the power supply reaches an output specification of 350 kV/60 mA.

  5. Unified optical distortion correction method for imaging systems using a concise geometrical transformation model

    NASA Astrophysics Data System (ADS)

    Chang, Shengqian; Liu, Siqi; Yuan, Fei; Zheng, Zhenrong

    2017-01-01

    Since optical distortion has been a big trouble for various kinds of imaging systems, finding a simple correction method with wide applications is of significant importance. In this paper, we propose a unified and simple correction method, performing well for both photographic and projective imaging systems. The basic idea is regarding the optical distortion as geometrical deformation between the object and image, without considering the specific features of an optical system. First of all, a calibration template is employed to establish the geometrical transformation model (GTM) for the distortion of a built optical system. Two alternative algorithms are given to estimate the GTM in algebraic form. The computation is very simple because no intrinsic parameters of the optical system are needed to establish the GTM. Besides, the errors introduced by the fabricating and assembling process can be eliminated. Then, the corrected image of the photographic system or the pre-distorted image of the projective systems can be obtained accordingly utilizing the GTM. Experiments are conducted to demonstrate the effectiveness of our method with wide applications.

  6. Novel Thermographic Method for Characterizing Transformation Temperatures of Thin-Film Shape Memory Alloys Aimed at Combinatorial Approach

    NASA Astrophysics Data System (ADS)

    Aono, Yuko; Sakurai, Junpei; Shimokohbe, Akira; Hata, Seiichi

    2011-06-01

    A novel characterization method for the two-way martensitic transformation temperatures of thin-film shape memory alloys (SMAs) is proposed. The method uses thermography to detect the transformation as a change in emissivity. The proposed method was demonstrated for a Ti-Pd-Ni thin-film SMA along with electrical resistivity monitoring. Definite emissivity changes were observed during both heating and cooling. Furthermore, the changes in emissivity corresponded to changes in the electrical resistivity. To confirm the validity of the proposed method, the results were compared with results obtained using conventional differential scanning calorimetry (DSC). The reverse-martensitic transformation temperature was found to agree with the results of the proposed method. However, the martensitic transformation temperature did not agree well. The reason for this disagreement is the differences in the definition of the transformation temperature and stress conditions used. We expect that the proposed method will enable high-throughput characterization of SMAs by a combinatorial approach in the future.

  7. Persistence of Free Plasmid DNA in Soil Monitored by Various Methods, Including a Transformation Assay

    PubMed Central

    Romanowski, Gerd; Lorenz, Michael G.; Sayler, Gary; Wackernagel, Wilfried

    1992-01-01

    The persistence and stability of free plasmid pUC8-ISP DNA introduced into 10-g samples of various soils and kept at 23°C were monitored over a period of 60 days. The soils were sampled at a plant science farm and included a loamy sand soil (no. 1), a clay soil (no. 2), and a silty clay soil (no. 3). Four different methods allowed monitoring of (i) the production of acid-soluble radioactive material from [3H]thymidine-labeled plasmid DNA, (ii) the decrease of hybridizing nucleotide sequences in slot blot analysis, (iii) the loss of plasmid integrity measured by Southern hybridization, and (iv) the decay of the biological activity as determined by transformation of Ca2+-treated Escherichia coli cells with the DNA extracted from soil. Acid-soluble material was not produced within the first 24 h but then increased to 45% (soil no. 1), 27% (soil no. 2), and 77% (soil no. 3) until the end of incubation. A quite parallel loss of material giving a slot blot hybridization signal was observed. Southern hybridization indicated that after 1 h in the soils, plasmid DNA was mostly in the form of circular and full-length linear molecules but that, depending on the soil type, after 2 to 5 days full-length plasmid molecules were hardly detectable. The transforming activity of plasmid DNA reextracted from the soils followed inactivation curves over 2 to 4 orders of magnitude and dropped below the detection limit after 10 days. The inactivation was slower in soil no. 2 (28.2-h half-life time of the transforming activity of a plasmid molecule) than in soils no. 3 (15.1 h) and no. 1 (9.1 h). The studies provide data on the persistence of free DNA molecules in natural bacterial soil habitats. The data suggest that plasmid DNA may persist long enough to be available for uptake by competent recipient cells in situ. Images PMID:16348772

  8. Numerical Methods for the Analysis of Power Transformer Tank Deformation and Rupture Due to Internal Arcing Faults.

    PubMed

    Yan, Chenguang; Hao, Zhiguo; Zhang, Song; Zhang, Baohui; Zheng, Tao

    2015-01-01

    Power transformer rupture and fire resulting from an arcing fault inside the tank usually leads to significant security risks and serious economic loss. In order to reveal the essence of tank deformation or explosion, this paper presents a 3-D numerical computational tool to simulate the structural dynamic behavior due to overpressure inside transformer tank. To illustrate the effectiveness of the proposed method, a 17.3 MJ and a 6.3 MJ arcing fault were simulated on a real full-scale 360MVA/220kV oil-immersed transformer model, respectively. By employing the finite element method, the transformer internal overpressure distribution, wave propagation and von-Mises stress were solved. The numerical results indicate that the increase of pressure and mechanical stress distribution are non-uniform and the stress tends to concentrate on connecting parts of the tank as the fault time evolves. Given this feature, it becomes possible to reduce the risk of transformer tank rupture through limiting the fault energy and enhancing the mechanical strength of the local stress concentrative areas. The theoretical model and numerical simulation method proposed in this paper can be used as a substitute for risky and costly field tests in fault overpressure analysis and tank mitigation design of transformers.

  9. Numerical Methods for the Analysis of Power Transformer Tank Deformation and Rupture Due to Internal Arcing Faults

    PubMed Central

    Yan, Chenguang; Hao, Zhiguo; Zhang, Song; Zhang, Baohui; Zheng, Tao

    2015-01-01

    Power transformer rupture and fire resulting from an arcing fault inside the tank usually leads to significant security risks and serious economic loss. In order to reveal the essence of tank deformation or explosion, this paper presents a 3-D numerical computational tool to simulate the structural dynamic behavior due to overpressure inside transformer tank. To illustrate the effectiveness of the proposed method, a 17.3MJ and a 6.3MJ arcing fault were simulated on a real full-scale 360MVA/220kV oil-immersed transformer model, respectively. By employing the finite element method, the transformer internal overpressure distribution, wave propagation and von-Mises stress were solved. The numerical results indicate that the increase of pressure and mechanical stress distribution are non-uniform and the stress tends to concentrate on connecting parts of the tank as the fault time evolves. Given this feature, it becomes possible to reduce the risk of transformer tank rupture through limiting the fault energy and enhancing the mechanical strength of the local stress concentrative areas. The theoretical model and numerical simulation method proposed in this paper can be used as a substitute for risky and costly field tests in fault overpressure analysis and tank mitigation design of transformers. PMID:26230392

  10. Application of the Fractional Fourier Transform and S-Method in Doppler Radar Tomography

    DTIC Science & Technology

    2010-08-01

    Tomography CPI - Coherent Processing Interval CW - Continuous Wave FFT - Fast Fourier Transform FrFT - Fractional Fourier Transform FT - Fourier Transform ISAR ...range profile of a rotating target from radar with a high sampling rate, which may be the case for radars with an ISAR capability. Alternatively, it may...modulated (LFM) audio signals from bats by Capus et al. in [11]. It has also been applied to the SAR and ISAR imaging problems of moving and

  11. Resonant frequency detection and adjustment method for a capacitive transducer with differential transformer bridge

    SciTech Connect

    Hu, M.; Bai, Y. Z. Zhou, Z. B. Li, Z. X.; Luo, J.

    2014-05-15

    The capacitive transducer with differential transformer bridge is widely used in ultra-sensitive space accelerometers due to their simple structure and high resolution. In this paper, the front-end electronics of an inductive-capacitive resonant bridge transducer is analyzed. The analysis result shows that the performance of this transducer depends upon the case that the AC pumping frequency operates at the resonance point of the inductive-capacitive bridge. The effect of possible mismatch between the AC pumping frequency and the actual resonant frequency is discussed, and the theoretical analysis indicates that the output voltage noise of the front-end electronics will deteriorate by a factor of about 3 due to either a 5% variation of the AC pumping frequency or a 10% variation of the tuning capacitance. A pre-scanning method to determine the actual resonant frequency is proposed followed by the adjustment of the operating frequency or the change of the tuning capacitance in order to maintain expected high resolution level. An experiment to verify the mismatching effect and the adjustment method is provided.

  12. Enzymatic transesterification monitored by an easy-to-use Fourier transform infrared spectroscopy method.

    PubMed

    Natalello, Antonino; Sasso, Francesco; Secundo, Francesco

    2013-01-01

    Transesterification of triglycerides with short chain alcohols is the key reaction in biodiesel production, in addition to other applications in chemical synthesis. However, it is crucial to optimize reaction conditions to make enzymatic transesterification a cost-effective and competitive process. In this work, a new, easy Fourier transform infrared (FTIR) spectroscopic approach for monitoring the transesterification reaction is reported and compared with a gas-chromatographic method. The concentration of the total methyl esters in the reaction mixture is determined from the peak intensity at ∼1435 cm(-1) in the second derivatives of the FTIR absorption spectra using a linear regression calibration. Interestingly, we found that the use of second derivatives allows an accurate determination of the methyl esters without the interference of free fatty acids. Moreover, information on substrate hydrolysis can be obtained within the same measurement by the infrared absorption at ∼1709 cm(-1) . We applied this approach to monitor methanolysis and hydrolysis reactions catalyzed by different commercial lipases, which displayed different sensitivities to methanol inhibition. Therefore, the FTIR approach reported in this work represents a rapid, inexpensive, and accurate method to monitor enzymatic transesterification, requiring very limited sample preparation and a simple statistical analysis of the spectroscopic data.

  13. Image restoration method based on Hilbert transform for full-field optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Na, Jihoon; Choi, Woo June; Choi, Eun Seo; Ryu, Seon Young; Lee, Byeong Ha

    2008-01-01

    A full-field optical coherence tomography (FF-OCT) system utilizing a simple but novel image restoration method suitable for a high-speed system is demonstrated. An en-face image is retrieved from only two phase-shifted interference fringe images through using the mathematical Hilbert transform. With a thermal light source, a high-resolution FF-OCT system having axial and transverse resolutions of 1 and 2.2 μm, respectively, was implemented. The feasibility of the proposed scheme is confirmed by presenting the obtained en-face images of biological samples such as a piece of garlic and a gold beetle. The proposed method is robust to the error in the amount of the phase shift and does not leave residual fringes. The use of just two interference images and the strong immunity to phase errors provide great advantages in the imaging speed and the system design flexibility of a high-speed high-resolution FF-OCT system.

  14. Applications of the conformal transformation method in studies of composed superconducting systems

    NASA Astrophysics Data System (ADS)

    Krzyzosiak, Mateusz; Gonczarek, Ryszard; Gonczarek, Adam; Jacak, Lucjan

    2016-12-01

    A framework for analytical studies of superconducting systems is presented and illustrated. The formalism, based on the conformal transformation of momentum space, allows one to study the effects of both the dispersion relation and the structure of the pairing interaction in two-dimensional anisotropic high- T c superconductors. In this method, the number of employed degrees of freedom coincides with the dimension of the momentum space, which is different compared to that in the standard Van Hove scenario with a single degree of freedom. A new function, the kernel of the density of states, is defined and its relation to the standard density of states is explained. The versatility of the method is illustrated by analyzing coexistence and competition between spin-singlet and spin-triplet order parameters in superconducting systems with a tight-binding-type dispersion relation and an anisotropic pairing potential. Phase diagrams of stable superconducting states in the coordinates η (the ratio of hopping parameters) and n (the carrier concentration) are presented and discussed. Moreover, the role of attractive and repulsive on-site interactions for the stability of the s-wave order parameter is explained.

  15. Method for calibrating a Fourier transform ion cyclotron resonance mass spectrometer

    DOEpatents

    Smith, Richard D.; Masselon, Christophe D.; Tolmachev, Aleksey

    2003-08-19

    A method for improving the calibration of a Fourier transform ion cyclotron resonance mass spectrometer wherein the frequency spectrum of a sample has been measured and the frequency (f) and intensity (I) of at least three species having known mass to charge (m/z) ratios and one specie having an unknown (m/z) ratio have been identified. The method uses the known (m/z) ratios, frequencies, and intensities at least three species to calculate coefficients A, B, and C, wherein the mass to charge ratio of a least one of the three species (m/z).sub.i is equal to ##EQU1## wherein f.sub.i is the detected frequency of the specie, G(I.sub.i) is a predetermined function of the intensity of the species, and Q is a predetermined exponent. Using the calculated values for A, B, and C, the mass to charge ratio of the unknown specie (m/z).sub.ii is calculated as the sum of ##EQU2## wherein f.sub.ii is the measured frequency of the unknown specie, and (I.sub.ii) is the measured intensity of the unknown specie.

  16. Sound quality recognition using optimal wavelet-packet transform and artificial neural network methods

    NASA Astrophysics Data System (ADS)

    Xing, Y. F.; Wang, Y. S.; Shi, L.; Guo, H.; Chen, H.

    2016-01-01

    According to the human perceptional characteristics, a method combined by the optimal wavelet-packet transform and artificial neural network, so-called OWPT-ANN model, for psychoacoustical recognition is presented. Comparisons of time-frequency analysis methods are performed, and an OWPT with 21 critical bands is designed for feature extraction of a sound, as is a three-layer back-propagation ANN for sound quality (SQ) recognition. Focusing on the loudness and sharpness, the OWPT-ANN model is applied on vehicle noises under different working conditions. Experimental verifications show that the OWPT can effectively transfer a sound into a time-varying energy pattern as that in the human auditory system. The errors of loudness and sharpness of vehicle noise from the OWPT-ANN are all less than 5%, which suggest a good accuracy of the OWPT-ANN model in SQ recognition. The proposed methodology might be regarded as a promising technique for signal processing in the human-hearing related fields in engineering.

  17. Recursion-transform method for computing resistance of the complex resistor network with three arbitrary boundaries

    NASA Astrophysics Data System (ADS)

    Tan, Zhi-Zhong

    2015-05-01

    We develop a general recursion-transform (R-T) method for a two-dimensional resistor network with a zero resistor boundary. As applications of the R-T method, we consider a significant example to illuminate the usefulness for calculating resistance of a rectangular m ×n resistor network with a null resistor and three arbitrary boundaries, a problem never solved before, since Green's function techniques and Laplacian matrix approaches are invalid in this case. Looking for the exact calculation of the resistance of a binary resistor network is important but difficult in the case of an arbitrary boundary since the boundary is like a wall or trap which affects the behavior of finite network. In this paper we obtain several general formulas of resistance between any two nodes in a nonregular m ×n resistor network in both finite and infinite cases. In particular, 12 special cases are given by reducing one of the general formulas to understand its applications and meanings, and an integral identity is found when we compare the equivalent resistance of two different structures of the same problem in a resistor network.

  18. Resonant frequency detection and adjustment method for a capacitive transducer with differential transformer bridge

    NASA Astrophysics Data System (ADS)

    Hu, M.; Bai, Y. Z.; Zhou, Z. B.; Li, Z. X.; Luo, J.

    2014-05-01

    The capacitive transducer with differential transformer bridge is widely used in ultra-sensitive space accelerometers due to their simple structure and high resolution. In this paper, the front-end electronics of an inductive-capacitive resonant bridge transducer is analyzed. The analysis result shows that the performance of this transducer depends upon the case that the AC pumping frequency operates at the resonance point of the inductive-capacitive bridge. The effect of possible mismatch between the AC pumping frequency and the actual resonant frequency is discussed, and the theoretical analysis indicates that the output voltage noise of the front-end electronics will deteriorate by a factor of about 3 due to either a 5% variation of the AC pumping frequency or a 10% variation of the tuning capacitance. A pre-scanning method to determine the actual resonant frequency is proposed followed by the adjustment of the operating frequency or the change of the tuning capacitance in order to maintain expected high resolution level. An experiment to verify the mismatching effect and the adjustment method is provided.

  19. Cross modulation method of transformation of the spatial coherence of pulsed laser radiation in a nonlinear medium

    SciTech Connect

    Kitsak, M A; Kitsak, A I

    2008-04-30

    The cross modulation method of transformation of the spatial coherence of low-power pulsed laser radiation in a nonlinear medium is proposed. The method is realised experimentally in a multimode optical fibre. The estimates of the degree of spatial coherence of radiation subjected to the phase cross modulation demonstrated the high efficiency of this radiation decorrelation mechanism. (control of laser radiation parameters)

  20. How to get exogenous DNA to cross the cell membrane of plants. Comment on “Physical methods for genetic transformation in plants” by Rivera et al.

    NASA Astrophysics Data System (ADS)

    Cruz Hernández, Andrés; Campos Guillén, Juan

    2012-09-01

    Physical methods for genetic transformation in plants. The most commonly applied methods in plant transformation include Agrobacterium infection and protoplast or microprojectile bombardment. A plant transformation system is a prerequisite for the development of a plant improvement program. The global area utilized for biotech crops increases every year.

  1. A simple method for optimising transformation of non-parametric data: an illustration by reference to cortisol assays.

    PubMed

    Clark, James E; Osborne, Jason W; Gallagher, Peter; Watson, Stuart

    2016-07-01

    Neuroendocrine data are typically positively skewed and rarely conform to the expectations of a Gaussian distribution. This can be a problem when attempting to analyse results within the framework of the general linear model, which relies on assumptions that residuals in the data are normally distributed. One frequently used method for handling violations of this assumption is to transform variables to bring residuals into closer alignment with assumptions (as residuals are not directly manipulated). This is often attempted through ad hoc traditional transformations such as square root, log and inverse. However, Box and Cox (Box & Cox, ) observed that these are all special cases of power transformations and proposed a more flexible method of transformation for researchers to optimise alignment with assumptions. The goal of this paper is to demonstrate the benefits of the infinitely flexible Box-Cox transformation on neuroendocrine data using syntax in spss. When applied to positively skewed data typical of neuroendocrine data, the majority (~2/3) of cases were brought into strict alignment with Gaussian distribution (i.e. a non-significant Shapiro-Wilks test). Those unable to meet this challenge showed substantial improvement in distributional properties. The biggest challenge was distributions with a high ratio of kurtosis to skewness. We discuss how these cases might be handled, and we highlight some of the broader issues associated with transformation. Copyright © 2016 John Wiley & Sons, Ltd.

  2. Theoretical Investigation of Thermo-Mechanical Behavior of Carbon Nanotube-Based Composites Using the Integral Transform Method

    NASA Technical Reports Server (NTRS)

    Pawloski, Janice S.

    2001-01-01

    This project uses the integral transform technique to model the problem of nanotube behavior as an axially symmetric system of shells. Assuming that the nanotube behavior can be described by the equations of elasticity, we seek a stress function x which satisfies the biharmonic equation: del(exp 4) chi = [partial deriv(r(exp 2)) + partial deriv(r) + partial deriv(z(exp 2))] chi = 0. The method of integral transformations is used to transform the differential equation. The symmetry with respect to the z-axis indicates that we only need to consider the sine transform of the stress function: X(bar)(r,zeta) = integral(from 0 to infinity) chi(r,z)sin(zeta,z) dz.

  3. Enhanced method to reconstruct mode shapes of continuous scanning measurements using the Hilbert Huang transform and the modal analysis method

    SciTech Connect

    Lee, Jongsuh; Hussain, Syed Hassaan; Wang, Semyung Park, Kyihwan

    2014-09-15

    Generally, it is time consuming to experimentally identify the operating deflection shape or mode shape of a structure. To overcome this problem, the Hilbert Huang transform (HHT) technique has been recently proposed. This technique is used to extract the mode shape from measurements that continuously measure the vibration of a region of interest within a structure using a non-contact laser sensor. In previous research regarding the HHT, two technical processes were needed to obtain the mode shape for each mode. The purpose of this study is to improve and complement our previous research, and for this purpose, a modal analysis approach is adapted without using the two technical processes to obtain an accurate un-damped impulse response of each mode for continuous scanning measurements. In addition, frequency response functions for each type of beam are derived, making it possible to make continuously scanned measurements along a straight profile. In this paper, the technical limitations and drawbacks of the damping compensation technique used in previous research are identified. In addition, the separation of resonant frequency (the Doppler effect) that occurs in continuous scanning measurements and the separation of damping phenomenon are also observed. The proposed method is quantitatively verified by comparing it with the results obtained from a conventional approach to estimate the mode shape with an impulse response.

  4. Enhanced method to reconstruct mode shapes of continuous scanning measurements using the Hilbert Huang transform and the modal analysis method.

    PubMed

    Lee, Jongsuh; Hussain, Syed Hassaan; Wang, Semyung; Park, Kyihwan

    2014-09-01

    Generally, it is time consuming to experimentally identify the operating deflection shape or mode shape of a structure. To overcome this problem, the Hilbert Huang transform (HHT) technique has been recently proposed. This technique is used to extract the mode shape from measurements that continuously measure the vibration of a region of interest within a structure using a non-contact laser sensor. In previous research regarding the HHT, two technical processes were needed to obtain the mode shape for each mode. The purpose of this study is to improve and complement our previous research, and for this purpose, a modal analysis approach is adapted without using the two technical processes to obtain an accurate un-damped impulse response of each mode for continuous scanning measurements. In addition, frequency response functions for each type of beam are derived, making it possible to make continuously scanned measurements along a straight profile. In this paper, the technical limitations and drawbacks of the damping compensation technique used in previous research are identified. In addition, the separation of resonant frequency (the Doppler effect) that occurs in continuous scanning measurements and the separation of damping phenomenon are also observed. The proposed method is quantitatively verified by comparing it with the results obtained from a conventional approach to estimate the mode shape with an impulse response.

  5. A transformation method for obtaining marker-free plants of a cross-pollinating and vegetatively propagated crop.

    PubMed

    de Vetten, Nick; Wolters, Anne-Marie; Raemakers, Krit; van der Meer, Ingrid; ter Stege, Renaldo; Heeres, Els; Heeres, Paul; Visser, Richard

    2003-04-01

    It is generally thought that transformation of plant cells using Agrobacterium tumefaciens occurs at a very low frequency. Therefore, selection marker genes are used to identify the rare plants that have taken up foreign DNA. Genes encoding antibiotic and herbicide resistance are widely used for this purpose in plant transformation. Over the past several years, consumer and environmental groups have expressed concern about the use of antibiotic- and herbicide-resistance genes from an ecological and food safety perspective. Although no scientific basis has been determined for these concerns, generating marker-free plants would certainly contribute to the public acceptance of transgenic crops. Several methods have been reported to create marker gene-free transformed plants, for example co-transformation, transposable elements, site-specific recombination, or intrachromosomal recombination. Not only are most of these systems time-consuming and inefficient, but they are also employed on the assumption that isolation of transformants without a selective marker gene is not feasible. Here we present a method that permits the identification of transgenic plants without the use of selectable markers. This strategy relies on the transformation of tissue explants or cells with a virulent A. tumefaciens strain and selection of transformed cells or shoots after PCR analysis. Incubation of potato explants with A. tumefaciens strain AGL0 resulted in transformed shoots at an efficiency of 1-5% of the harvested shoots, depending on the potato genotype used. Because this system does not require genetic segregation or site-specific DNA-deletion systems to remove marker genes, it may provide a reliable and efficient tool for generating transgenic plants for commercial use, especially in vegetatively propagated species like potato and cassava.

  6. Quantitative Estimation of Trace Chemicals in Industrial Effluents with the Sticklet Transform Method

    SciTech Connect

    Mehta, N C; Scharlemann, E T; Stevens, C G

    2001-04-02

    Application of a novel transform operator, the Sticklet transform, to the quantitative estimation of trace chemicals in industrial effluent plumes is reported. The sticklet transform is a superset of the well-known derivative operator and the Haar wavelet, and is characterized by independently adjustable lobe width and separation. Computer simulations demonstrate that they can make accurate and robust concentration estimates of multiple chemical species in industrial effluent plumes in the presence of strong clutter background, interferent chemicals and random noise. In this paper they address the application of the sticklet transform in estimating chemical concentrations in effluent plumes in the presence of atmospheric transmission effects. They show that this transform retains the ability to yield accurate estimates using on-plume/off-plume measurements that represent atmospheric differentials up to 10% of the full atmospheric attenuation.

  7. Detecting the quality of glycerol monolaurate: a method for using Fourier transform infrared spectroscopy with wavelet transform and modified uninformative variable elimination.

    PubMed

    Chen, Xiaojing; Wu, Di; He, Yong; Liu, Shou

    2009-04-06

    Glycerol monolaurate (GML) products contain many impurities, such as lauric acid and glucerol. The GML content is an important quality indicator for GML production. A hybrid variable selection algorithm, which is a combination of wavelet transform (WT) technology and modified uninformative variable eliminate (MUVE) method, was proposed to extract useful information from Fourier transform infrared (FT-IR) transmission spectroscopy for the determination of GML content. FT-IR spectra data were compressed by WT first; the irrelevant variables in the compressed wavelet coefficients were eliminated by MUVE. In the MUVE process, simulated annealing (SA) algorithm was employed to search the optimal cutoff threshold. After the WT-MUVE process, variables for the calibration model were reduced from 7366 to 163. Finally, the retained variables were employed as inputs of partial least squares (PLS) model to build the calibration model. For the prediction set, the correlation coefficient (r) of 0.9910 and root mean square error of prediction (RMSEP) of 4.8617 were obtained. The prediction result was better than the PLS model with full-spectra data. It was indicated that proposed WT-MUVE method could not only make the prediction more accurate, but also make the calibration model more parsimonious. Furthermore, the reconstructed spectra represented the projection of the selected wavelet coefficients into the original domain, affording the chemical interpretation of the predicted results. It is concluded that the FT-IR transmission spectroscopy technique with the proposed method is promising for the fast detection of GML content.

  8. A protocol for sonication-assisted Agrobacterium rhizogenes-mediated transformation of haploid and diploid sugar beet (Beta vulgaris L.) explants.

    PubMed

    Klimek-Chodacka, Magdalena; Baranski, Rafal

    2014-01-01

    Hairy root cultures obtained after Agrobacterium rhizogenes-mediated genetic transformation can serve as a model system for studying plant metabolism and physiology, or can be utilized for the production of secondary metabolites. So far no efficient protocol of hairy root development in sugar beet has been publically released. In this work, two A. rhizogenes strains (A4T and LBA1334) carrying a binary vector pBIN-m-gfp5-ER or pCAMBIA1301 possessing gfp and uidA reporter genes were used to transform petiole explants of haploid and diploid sugar beet genotypes. Five treatment combinations of sonicated-assisted Agrobacterium-mediated transformation were compared. Hairy roots appeared on 0% to 54% of explants depending on the treatment combination used. The highest frequency was achieved when explants of a diploid genotype were sonicated for 15 s in the inoculum containing A. rhizogenes of OD600=0.5 and then co-cultured for three days. Using the same treatment combinations the explants of haploid genotypes developed hairy roots with the frequency ranging from 10% to 36%. Transformation efficiency was independent on the bacterial strain used. The results indicate that haploid sugar beet explants are amenable to transformation using A. rhizogenes, and that the efficiency of that process can be increased by applying short ultrasound treatment.

  9. Transformation of Lettuce with rol ABC Genes: Extracts Show Enhanced Antioxidant, Analgesic, Anti-Inflammatory, Antidepressant, and Anticoagulant Activities in Rats.

    PubMed

    Ismail, Hammad; Dilshad, Erum; Waheed, Mohammad Tahir; Mirza, Bushra

    2017-03-01

    Lettuce is an edible crop that is well known for dietary and antioxidant benefits. The present study was conducted to investigate the effects of rol ABC genes on antioxidant and medicinal potential of lettuce by Agrobacterium-mediated transformation. Transgene integration and expression was confirmed through PCR and real-time RT-PCR, respectively. The transformed plants showed 91-102 % increase in total phenolic contents and 53-65 % increase in total flavonoid contents compared to untransformed plants. Total antioxidant capacity and total reducing power increased up to 112 and 133 % in transformed plants, respectively. Results of DPPH assay showed maximum 51 % increase, and lipid peroxidation assay exhibited 20 % increase in antioxidant activity of transformed plants compared to controls. Different in vivo assays were carried out in rats. The transgenic plants showed up to 80 % inhibition in both hot plate analgesic assay and carrageenan-induced hind paw edema test, while untransformed plants showed only 45 % inhibition. Antidepressant and anticoagulant potential of transformed plants was also significantly enhanced compared to untransformed plants. Taken together, the present work highlights the use of rol genes to enhance the secondary metabolite production in lettuce and improve its analgesic, anti-inflammatory, antidepressant, and anticoagulatory properties.

  10. Multistep generalized transformation method applied to solving equations of discrete and continuous time-fractional enzyme kinetics

    NASA Astrophysics Data System (ADS)

    Vosika, Z.; Mitić, V. V.; Vasić, A.; Lazović, G.; Matija, L.; Kocić, Lj. M.

    2017-03-01

    In this paper, Caputo based Michaelis-Menten kinetic model based on Time Scale Calculus (TSC) is proposed. The main reason for its consideration is a study of tumor cells population growth dynamics. In the particular case discrete-continuous time kinetics, Michaelis-Menten model is numerically treated, using a new algorithm proposed by authors, called multistep generalized difference transformation method (MSGDETM). In addition numerical simulations are performed and is shown that it represents the upgrade of the multi-step variant of generalized differential transformation method (MSGDTM). A possible conditions for its further development are discussed and possible experimental verification is described.

  11. Effectiveness of dereverberation, feature transformation, discriminative training methods, and system combination approach for various reverberant environments

    NASA Astrophysics Data System (ADS)

    Tachioka, Yuuki; Narita, Tomohiro; Watanabe, Shinji

    2015-12-01

    The recently released REverberant Voice Enhancement and Recognition Benchmark (REVERB) challenge includes a reverberant automatic speech recognition (ASR) task. This paper describes our proposed system based on multi-channel speech enhancement preprocessing and state-of-the-art ASR techniques. For preprocessing, we propose a single-channel dereverberation method with reverberation time estimation, which is combined with multichannel beamforming that enhances direct sound compared with the reflected sound. In addition, this paper also focuses on state-of-the-art ASR techniques such as discriminative training of acoustic models including the Gaussian mixture model, subspace Gaussian mixture model, and deep neural networks, as well as various feature transformation techniques. Although, for the REVERB challenge, it is necessary to handle various acoustic environments, a single ASR system tends to be overly tuned for a specific environment, which degrades the performance in the mismatch environments. To overcome this mismatch problem with a single ASR system, we use a system combination approach using multiple ASR systems with different features and different model types because a combination of various systems that have different error patterns is beneficial. In particular, we use our discriminative training technique for system combination that achieves better generalization by making systems complementary with the modified discriminative criteria. Experiments show the effectiveness of these approaches, reaching 6.76 and 18.60 % word error rates on the REVERB simulated and real test sets. These are 68.8 and 61.5 % relative improvements over the baseline.

  12. Light calibration and quality assessment methods for Reflectance Transformation Imaging applied to artworks' analysis

    NASA Astrophysics Data System (ADS)

    Giachetti, A.; Daffara, C.; Reghelin, C.; Gobbetti, E.; Pintus, R.

    2015-06-01

    In this paper we analyze some problems related to the acquisition of multiple illumination images for Polynomial Texture Maps (PTM) or generic Reflectance Transform Imaging (RTI). We show that intensity and directionality nonuniformity can be a relevant issue when acquiring manual sets of images with the standard highlight-based setup both using a flash lamp and a LED light. To maintain a cheap and flexible acquisition setup that can be used on field and by non-experienced users we propose to use a dynamic calibration and correction of the lights based on multiple intensity and direction estimation around the imaged object during the acquisition. Preliminary tests on the results obtained have been performed by acquiring a specifically designed 3D printed pattern in order to see the accuracy of the acquisition obtained both for spatial discrimination of small structures and normal estimation, and on samples of different types of paper in order to evaluate material discrimination. We plan to design and build from our analysis and from the tools developed and under development a set of novel procedures and guidelines that can be used to turn the cheap and common RTI acquisition setup from a simple way to enrich object visualization into a powerful method for extracting quantitative characterization both of surface geometry and of reflective properties of different materials. These results could have relevant applications in the Cultural Heritage domain, in order to recognize different materials used in paintings or investigate the ageing status of artifacts' surface.

  13. A method to correct sampling ghosts in historic near-infrared Fourier Transform Spectrometer (FTS) measurements

    NASA Astrophysics Data System (ADS)

    Dohe, S.; Sherlock, V.; Hase, F.; Gisi, M.; Robinson, J.; Sepúlveda, E.; Schneider, M.; Blumenstock, T.

    2013-04-01

    The Total Carbon Column Observing Network (TCCON) has been established to provide ground-based remote sensing measurements of the column-average dry air mole fractions of key greenhouse gases. To ensure the network wide consistency, biases between Fourier Transform spectrometers at different sites have to be well controlled. In this study we investigate a fundamental correction scheme for errors in the sampling of the interferogram. This is a two-step procedure in which the laser sampling error (LSE) is quantified using a subset of suitable interferograms and then used to resample all the interferograms in the timeseries. Timeseries of measurements acquired at the TCCON sites Izaña and Lauder are used to demonstrate the method. At both sites the sampling error histories show changes in LSE due to instrument interventions. Estimated LSE are in good agreement with sampling errors inferred from lamp measurements of the ghost to parent ratio (Lauder). The LSE introduce retrieval biases which are minimised when the interferograms are resampled. The original timeseries of Xair and XCO2 at both sites show discrepancies of 0.2-0.5% due to changes in the LSE associated with instrument interventions or changes in the measurement sample rate. After resampling discrepancies are reduced to 0.1% at Lauder and 0.2% at Izaña. In the latter case, coincident changes in interferometer alignment may also contribute to the residual difference.

  14. A method to correct sampling ghosts in historic near-infrared Fourier transform spectrometer (FTS) measurements

    NASA Astrophysics Data System (ADS)

    Dohe, S.; Sherlock, V.; Hase, F.; Gisi, M.; Robinson, J.; Sepúlveda, E.; Schneider, M.; Blumenstock, T.

    2013-08-01

    The Total Carbon Column Observing Network (TCCON) has been established to provide ground-based remote sensing measurements of the column-averaged dry air mole fractions (DMF) of key greenhouse gases. To ensure network-wide consistency, biases between Fourier transform spectrometers at different sites have to be well controlled. Errors in interferogram sampling can introduce significant biases in retrievals. In this study we investigate a two-step scheme to correct these errors. In the first step the laser sampling error (LSE) is estimated by determining the sampling shift which minimises the magnitude of the signal intensity in selected, fully absorbed regions of the solar spectrum. The LSE is estimated for every day with measurements which meet certain selection criteria to derive the site-specific time series of the LSEs. In the second step, this sequence of LSEs is used to resample all the interferograms acquired at the site, and hence correct the sampling errors. Measurements acquired at the Izaña and Lauder TCCON sites are used to demonstrate the method. At both sites the sampling error histories show changes in LSE due to instrument interventions (e.g. realignment). Estimated LSEs are in good agreement with sampling errors inferred from the ratio of primary and ghost spectral signatures in optically bandpass-limited tungsten lamp spectra acquired at Lauder. The original time series of Xair and XCO2 (XY: column-averaged DMF of the target gas Y) at both sites show discrepancies of 0.2-0.5% due to changes in the LSE associated with instrument interventions or changes in the measurement sample rate. After resampling, discrepancies are reduced to 0.1% or less at Lauder and 0.2% at Izaña. In the latter case, coincident changes in interferometer alignment may also have contributed to the residual difference. In the future the proposed method will be used to correct historical spectra at all TCCON sites.

  15. Analytical solutions for solute transport in groundwater and riverine flow using Green's Function Method and pertinent coordinate transformation method

    NASA Astrophysics Data System (ADS)

    Sanskrityayn, Abhishek; Suk, Heejun; Kumar, Naveen

    2017-04-01

    In this study, analytical solutions of one-dimensional pollutant transport originating from instantaneous and continuous point sources were developed in groundwater and riverine flow using both Green's Function Method (GFM) and pertinent coordinate transformation method. Dispersion coefficient and flow velocity are considered spatially and temporally dependent. The spatial dependence of the velocity is linear, non-homogeneous and that of dispersion coefficient is square of that of velocity, while the temporal dependence is considered linear, exponentially and asymptotically decelerating and accelerating. Our proposed analytical solutions are derived for three different situations depending on variations of dispersion coefficient and velocity, respectively which can represent real physical processes occurring in groundwater and riverine systems. First case refers to steady solute transport situation in steady flow in which dispersion coefficient and velocity are only spatially dependent. The second case represents transient solute transport in steady flow in which dispersion coefficient is spatially and temporally dependent while the velocity is spatially dependent. Finally, the third case indicates transient solute transport in unsteady flow in which both dispersion coefficient and velocity are spatially and temporally dependent. The present paper demonstrates the concentration distribution behavior from a point source in realistically occurring flow domains of hydrological systems including groundwater and riverine water in which the dispersivity of pollutant's mass is affected by heterogeneity of the medium as well as by other factors like velocity fluctuations, while velocity is influenced by water table slope and recharge rate. Such capabilities give the proposed method's superiority about application of various hydrological problems to be solved over other previously existing analytical solutions. Especially, to author's knowledge, any other solution doesn

  16. Finite element Fourier and Abbe transform methods for generalization of aperture function and geometry in Fraunhofer diffraction theory

    SciTech Connect

    Kraus, H.G. )

    1991-08-01

    This paper discusses methods for calculating Fraunhofer intensity fields resulting from diffraction through one- and two-dimensional apertures are presented. These methods are based on the geometric concept of finite elements and on Fourier and Abbe transforms. The geometry of the two-dimensional diffracting aperture(s) is based on biquadratic isoparametric elements, which are used to define aperture(s) of complex geometry. These elements are also used to build complex amplitude and phase functions across the aperture(s) which may be of continuous or discontinuous form. The transform integrals are accurately and efficiently integrated numerically using Gaussian quadrature. The power of these methods is most evident in two dimensions, where several examples are presented which include secondary obstructions, straight and curved secondary spider supports, multiple-mirror arrays, synthetic aperture arrays, segmented mirrors, apertures covered by screens, apodization, and phase plates. Typically, the finite element Abbe transform method results in significant gains in computational efficiency over the finite element Fourier transform method, but is also subject to some loss in generality.

  17. Method for distinguishing normal and transformed cells using G1 kinase inhibitors

    DOEpatents

    Crissman, Harry A.; Gadbois, Donna M.; Tobey, Robert A.; Bradbury, E. Morton

    1993-01-01

    A G.sub.1 phase kinase inhibitor is applied in a low concentration to a population of normal and transformed mammalian cells. The concentration of G.sub.1 phase kinase inhibitor is selected to reversibly arrest normal mammalian cells in the G.sub.1 cell cycle without arresting growth of transformed cells. The transformed cells may then be selectively identified and/or cloned for research or diagnostic purposes. The transformed cells may also be selectively killed by therapeutic agents that do not affect normal cells in the G.sub.1 phase, suggesting that such G.sub.1 phase kinase inhibitors may form an effective adjuvant for use with chemotherapeutic agents in cancer therapy for optimizing the killing dose of chemotherapeutic agents while minimizing undesirable side effects on normal cells.

  18. Method for distinguishing normal and transformed cells using G1 kinase inhibitors

    DOEpatents

    Crissman, H.A.; Gadbois, D.M.; Tobey, R.A.; Bradbury, E.M.

    1993-02-09

    A G[sub 1] phase kinase inhibitor is applied in a low concentration to a population of normal and transformed mammalian cells. The concentration of G[sub 1] phase kinase inhibitor is selected to reversibly arrest normal mammalian cells in the G[sub 1] cell cycle without arresting growth of transformed cells. The transformed cells may then be selectively identified and/or cloned for research or diagnostic purposes. The transformed cells may also be selectively killed by therapeutic agents that do not affect normal cells in the G[sub 1] phase, suggesting that such G[sub 1] phase kinase inhibitors may form an effective adjuvant for use with chemotherapeutic agents in cancer therapy for optimizing the killing dose of chemotherapeutic agents while minimizing undesirable side effects on normal cells.

  19. Method for distinguishing normal and transformed cells using G1 kinase inhibitors

    SciTech Connect

    Crissman, H.A.; Gadbois, D.M.; Tobey, R.A.; Bradbury, E.M.

    1991-12-31

    A G{sub 1} phase kinase inhibitor is applied in a low concentration to a population of normal and transformed mammalian cells. The concentration of G{sub 1} phase kinase inhibitor is selected to reversibly arrest normal mammalian cells in the G{sub 1} cell cycle without arresting growth of transformed cells. The transformed cells may then be selectively identified and/or cloned for research or diagnostic purposes. The transformed cells may also be selectively killed by therapeutic agents that do not affect normal cells in the G{sub 1} phase, suggesting that such G{sub 1} phase kinase inhibitors may form an effective adjuvant for use with chemotherapeutic agents in cancer therapy for optimizing the killing dose of chemotherapeutic agents while minimizing undesirable side effects on normal cells.

  20. General method of the relativistic Foldy-Wouthuysen transformation and proof of validity of the Foldy-Wouthuysen Hamiltonian

    NASA Astrophysics Data System (ADS)

    Silenko, Alexander J.

    2015-02-01

    A general method of the Foldy-Wouthuysen transformation is developed. This method is applicable to relativistic particles with any spin in arbitrarily strong external fields. It can be used when the de Broglie wavelength is much smaller than the characteristic distance. Contrary to previously developed relativistic methods, the present method satisfies the condition of the exact Foldy-Wouthuysen transformation and is well substantiated. The derived relativistic Foldy-Wouthuysen Hamiltonian is expanded in powers of the Planck constant. In this expansion, terms proportional to the zero and first powers are determined exactly in accordance with the above condition, and terms proportional to higher powers are not specified. The obtained result agrees with the corresponding formula for the Foldy-Wouthuysen Hamiltonian previously deduced by an iterative relativistic method and proves the validity of results obtained with this formula.

  1. Novel image fusion method based on adaptive pulse coupled neural network and discrete multi-parameter fractional random transform

    NASA Astrophysics Data System (ADS)

    Lang, Jun; Hao, Zhengchao

    2014-01-01

    In this paper, we first propose the discrete multi-parameter fractional random transform (DMPFRNT), which can make the spectrum distributed randomly and uniformly. Then we introduce this new spectrum transform into the image fusion field and present a new approach for the remote sensing image fusion, which utilizes both adaptive pulse coupled neural network (PCNN) and the discrete multi-parameter fractional random transform in order to meet the requirements of both high spatial resolution and low spectral distortion. In the proposed scheme, the multi-spectral (MS) and panchromatic (Pan) images are converted into the discrete multi-parameter fractional random transform domains, respectively. In DMPFRNT spectrum domain, high amplitude spectrum (HAS) and low amplitude spectrum (LAS) components carry different informations of original images. We take full advantage of the synchronization pulse issuance characteristics of PCNN to extract the HAS and LAS components properly, and give us the PCNN ignition mapping images which can be used to determine the fusion parameters. In the fusion process, local standard deviation of the amplitude spectrum is chosen as the link strength of pulse coupled neural network. Numerical simulations are performed to demonstrate that the proposed method is more reliable and superior than several existing methods based on Hue Saturation Intensity representation, Principal Component Analysis, the discrete fractional random transform etc.

  2. Selective Weighted Least Squares Method for Fourier Transform Infrared Quantitative Analysis.

    PubMed

    Wang, Xin; Li, Yan; Wei, Haoyun; Chen, Xia

    2016-10-26

    Classical least squares (CLS) regression is a popular multivariate statistical method used frequently for quantitative analysis using Fourier transform infrared (FT-IR) spectrometry. Classical least squares provides the best unbiased estimator for uncorrelated residual errors with zero mean and equal variance. However, the noise in FT-IR spectra, which accounts for a large portion of the residual errors, is heteroscedastic. Thus, if this noise with zero mean dominates in the residual errors, the weighted least squares (WLS) regression method described in this paper is a better estimator than CLS. However, if bias errors, such as the residual baseline error, are significant, WLS may perform worse than CLS. In this paper, we compare the effect of noise and bias error in using CLS and WLS in quantitative analysis. Results indicated that for wavenumbers with low absorbance, the bias error significantly affected the error, such that the performance of CLS is better than that of WLS. However, for wavenumbers with high absorbance, the noise significantly affected the error, and WLS proves to be better than CLS. Thus, we propose a selective weighted least squares (SWLS) regression that processes data with different wavenumbers using either CLS or WLS based on a selection criterion, i.e., lower or higher than an absorbance threshold. The effects of various factors on the optimal threshold value (OTV) for SWLS have been studied through numerical simulations. These studies reported that: (1) the concentration and the analyte type had minimal effect on OTV; and (2) the major factor that influences OTV is the ratio between the bias error and the standard deviation of the noise. The last part of this paper is dedicated to quantitative analysis of methane gas spectra, and methane/toluene mixtures gas spectra as measured using FT-IR spectrometry and CLS, WLS, and SWLS. The standard error of prediction (SEP), bias of prediction (bias), and the residual sum of squares of the errors

  3. A Mixed-Methods Study of the Transformation Model for Rapid Improvement of Low Achieving Rural Schools

    ERIC Educational Resources Information Center

    Atkinson Duina, Angela

    2013-01-01

    New regulations attached to ARRA funding of federal School Improvement Fund grants aimed at producing rapid turnaround of low performing schools were highly criticized as unsuitable for rural schools. This mixed-methods study looked at the implementation of the School Improvement Fund Transformation Model in two rural Maine high schools during the…

  4. Evolution of heavy quark distribution function on quark-gluon plasma: Using the Iterative Laplace Transform Method

    NASA Astrophysics Data System (ADS)

    Mehrabi Pari, Sharareh; Javidan, Kurosh; Taghavi Shahri, Fatemeh

    2016-05-01

    The "Laplace Transform Method" is used to solve the Fokker-Plank equation for finding the time evolution of the heavy quarks distribution functions such as charm and bottom in quark gluon plasma. These solutions will lead us to calculation of nuclear suppression factor RAA. The results have good agreement with available experiment data from the PHENIX collaboration.

  5. 10 CFR Appendix A to Subpart K of... - Uniform Test Method for Measuring the Energy Consumption of Distribution Transformers

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Measured quantity Test systemaccuracy Power Losses ± 3.0% Voltage ± 0.5% Current ± 0.5% Resistance ± 0.5... establish the winding temperature at the time of the winding resistance measurement. (b) Measure the direct current resistance (Rdc) of transformer windings by one of the methods outlined in section 3.3....

  6. 10 CFR Appendix A to Subpart K of... - Uniform Test Method for Measuring the Energy Consumption of Distribution Transformers

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Measured quantity Test systemaccuracy Power Losses ±3.0% Voltage ±0.5% Current ±0.5% Resistance ±0.5... establish the winding temperature at the time of the winding resistance measurement. (b) Measure the direct current resistance (Rdc) of transformer windings by one of the methods outlined in section 3.3....

  7. Potato (Solanum tuberosum L.).

    PubMed

    Chetty, Venkateswari J; Narváez-Vásquez, Javier; Orozco-Cárdenas, Martha L

    2015-01-01

    Agrobacterium-mediated transformation is the most common method for the incorporation of foreign genes into the genome of potato as well as many other species in the Solanaceae family. This chapter describes protocols for the genetic transformation of three species of potato: Solanum tuberosum subsp. tuberosum (Desiréé), S. tuberosum subsp. andigenum (Blue potato), and S. tuberosum subsp. andigena using internodal segments as explants.

  8. A comparative analysis of Painleve, Lax pair, and similarity transformation methods in obtaining the integrability conditions of nonlinear Schroedinger equations

    SciTech Connect

    Al Khawaja, U.

    2010-05-15

    We derive the integrability conditions of nonautonomous nonlinear Schroedinger equations using the Lax pair and similarity transformation methods. We present a comparative analysis of these integrability conditions with those of the Painleve method. We show that while the Painleve integrability conditions restrict the dispersion, nonlinearity, and dissipation/gain coefficients to be space independent and the external potential to be only a quadratic function of position, the Lax Pair and the similarity transformation methods allow for space-dependent coefficients and an external potential that is not restricted to the quadratic form. The integrability conditions of the Painleve method are retrieved as a special case of our general integrability conditions. We also derive the integrability conditions of nonautonomous nonlinear Schroedinger equations for two- and three-spacial dimensions.

  9. Homotopy perturbation method with Laplace Transform (LT-HPM) for solving Lane-Emden type differential equations (LETDEs).

    PubMed

    Tripathi, Rajnee; Mishra, Hradyesh Kumar

    2016-01-01

    In this communication, we describe the Homotopy Perturbation Method with Laplace Transform (LT-HPM), which is used to solve the Lane-Emden type differential equations. It's very difficult to solve numerically the Lane-Emden types of the differential equation. Here we implemented this method for two linear homogeneous, two linear nonhomogeneous, and four nonlinear homogeneous Lane-Emden type differential equations and use their appropriate comparisons with exact solutions. In the current study, some examples are better than other existing methods with their nearer results in the form of power series. The Laplace transform used to accelerate the convergence of power series and the results are shown in the tables and graphs which have good agreement with the other existing method in the literature. The results show that LT-HPM is very effective and easy to implement.

  10. Numerical solutions and solitary wave solutions of fractional KDV equations using modified fractional reduced differential transform method

    NASA Astrophysics Data System (ADS)

    Saha Ray, S.

    2013-12-01

    In this paper, the modified fractional reduced differential transform method (MFRDTM) has been proposed and it is implemented for solving fractional KdV (Korteweg-de Vries) equations. The fractional derivatives are described in the Caputo sense. In this paper, the reduced differential transform method is modified to be easily employed to solve wide kinds of nonlinear fractional differential equations. In this new approach, the nonlinear term is replaced by its Adomian polynomials. Thus the nonlinear initial-value problem can be easily solved with less computational effort. In order to show the power and effectiveness of the present modified method and to illustrate the pertinent features of the solutions, several fractional KdV equations with different types of nonlinearities are considered. The results reveal that the proposed method is very effective and simple for obtaining approximate solutions of fractional KdV equations.

  11. Development of a transgenic hairy root system in jute (Corchorus capsularis L.) with gusA reporter gene through Agrobacterium rhizogenes mediated co-transformation.

    PubMed

    Chattopadhyay, Tirthartha; Roy, Sheuli; Mitra, Adinpunya; Maiti, Mrinal K

    2011-04-01

    Transgenic hairy root system is important in several recalcitrant plants, where Agrobacterium tumefaciens-mediated plant transformation and generation of transgenic plants are problematic. Jute (Corchorus spp.), the major fibre crop in Indian subcontinent, is one of those recalcitrant plants where in vitro tissue culture has provided a little success, and hence, Agrobacterium-mediated genetic transformation remains to be a challenging proposition in this crop. In the present work, a system of transgenic hairy roots in Corchorus capsularis L. has been developed through genetic transformation by Agrobacterium rhizogenes harbouring two plasmids, i.e. the natural Ri plasmid and a recombinant binary vector derived from the disarmed Ti plasmid of A. tumefaciens. Our findings indicate that the system is relatively easy to establish and reproducible. Molecular analysis of the independent lines of transgenic hairy roots revealed the transfer of relevant transgenes from both the T-DNA parts into the plant genome, indicating the co-transformation nature of the event. High level expression and activity of the gusA reporter gene advocate that the transgenic hairy root system, thus developed, could be applicable as gene expression system in general and for root functional genomics in particular. Furthermore, these transgenic hairy roots can be used in future as explants for plantlet regeneration to obtain stable transgenic jute plants.

  12. Fourier transform infrared spectroscopy as a method to study lipid accumulation in oleaginous yeasts

    PubMed Central

    2014-01-01

    Background Oleaginous microorganisms, such as different yeast and algal species, can represent a sustainable alternative to plant oil for the production of biodiesel. They can accumulate fatty acids (FA) up to 70% of their dry weight with a predominance of (mono)unsaturated species, similarly to what plants do, but differently from animals. In addition, their growth is not in competition either with food, feed crops, or with agricultural land. Despite these advantages, the exploitation of the single cell oil system is still at an early developmental stage. Cultivation mode and conditions, as well as lipid extraction technologies, represent the main limitations. The monitoring of lipid accumulation in oleaginous microorganisms is consequently crucial to develop and validate new approaches, but at present the majority of the available techniques is time consuming, invasive and, when relying on lipid extraction, can be affected by FA degradation. Results In this work the fatty acid accumulation of the oleaginous yeasts Cryptococcus curvatus and Rhodosporidium toruloides and of the non-oleaginous yeast Saccharomyces cerevisiae (as a negative control) was monitored in situ by Fourier Transform Infrared Spectroscopy (FTIR). Indeed, this spectroscopic tool can provide complementary information to those obtained by classical techniques, such as microscopy, flow cytometry and gas chromatography. As shown in this work, through the analysis of the absorption spectra of intact oleaginous microorganisms it is possible not only to monitor the progression of FA accumulation but also to identify the most represented classes of the produced lipids. Conclusions Here we propose FTIR microspectroscopy - supported by multivariate analysis - as a fast, reliable and non invasive method to monitor and analyze FA accumulation in intact oleaginous yeasts. The results obtained by the FTIR approach were in agreement with those obtained by the other classical methods like flow cytometry and

  13. A combined method for triplex pump fault diagnosis based on wavelet transform, fuzzy logic and neuro-networks

    NASA Astrophysics Data System (ADS)

    Kong, Fansen; Chen, Ruheng

    2004-01-01

    A new combined method based on wavelet transformation, fuzzy logic and neuro-networks is proposed for fault diagnosis of a triplex. The failure characteristics of the fluid- and dynamic-end can be divided into wavelet transform in different scales at the same time (in: Jun Zhu et al. (Eds.), Proceedings of an International Conference on Condition Monitoring. National Defense Industry Press, Beijing, 1997, pp. 271-275). Therefore, the characteristic variables can be constructed making use of the coefficients of Edgeworth asymptotic spectrum expansion formula and fuzzified to train the neuro-network to identify the faults of fluid- and dynamic-end of triplex pump in fuzzy domain. Tests indicate that the information of wavelet transformation in scale 2 is related to the meshing state of the gear and the information in scales 4 and 5 is related to the running state of fluid-end. Good agreement between analytical and experimental results has been obtained.

  14. A Method of Speech Periodicity Enhancement Using Transform-domain Signal Decomposition.

    PubMed

    Huang, Huang; Lee, Tan; Kleijn, W Bastiaan; Kong, Ying-Yee

    2015-03-01

    Periodicity is an important property of speech signals. It is the basis of the signal's fundamental frequency and the pitch of voice, which is crucial to speech communication. This paper presents a novel framework of periodicity enhancement for noisy speech. The enhancement is applied to the linear prediction residual of speech. The residual signal goes through a constant-pitch time warping process and two sequential lapped-frequency transforms, by which the periodic component is concentrated in certain transform coefficients. By emphasizing the respective transform coefficients, periodicity enhancement of noisy residual signal is achieved. The enhanced residual signal and estimated linear prediction filter parameters are used to synthesize the output speech. An adaptive algorithm is proposed for adjusting the weights for the periodic and aperiodic components. Effectiveness of the proposed approach is demonstrated via experimental evaluation. It is observed that harmonic structure of the original speech could be properly restored to improve the perceptual quality of enhanced speech.

  15. An asymmetric color image encryption method by using deduced gyrator transform

    NASA Astrophysics Data System (ADS)

    Yao, Lili; Yuan, Caojin; Qiang, Junjie; Feng, Shaotong; Nie, Shouping

    2017-02-01

    An encryption algorithm is proposed by using the properties of deduced gyrator transform (GT). After being transformed by the GT algorithm and multiplied by a phase distribution p*, the spectrum modulus of the input image is considered to be the encrypted image by further performing Fourier transformation. To resist the attack from iterative phase retrieval, the red, green and blue components of the input image is modulated by a random phase mask and then combined using convolution. The encryption result is real-valued, which is convenient for display, transmission and storage. In the decryption process, the three original color components can be recovered with decryption keys which are different from the encryption keys. An optoelectronic hybrid system for the encryption process is also presented. Computer simulations are presented to demonstrate its performance, and the security of the proposed system is analyzed as well.

  16. Stable transformation of Theobroma cacao L. and influence of matrix attachment regions on GFP expression.

    PubMed

    Maximova, S; Miller, C; Antúnez de Mayolo, G; Pishak, S; Young, A; Guiltinan, M J

    2003-06-01

    We describe a protocol for Agrobacterium-mediated genetic transformation of Theobroma cacao L. using cotyledonary explants from primary somatic embryos (SEs) and A. tumefaciens strain AGL1. Transgenic plants carrying the visible marker, gene green fluorescent protein ( EGFP), the selectable marker gene neomycin phosphotransferase II ( NPTII), the class I chitinase gene from cacao ( Chi), and tobacco nuclear matrix attachment regions (MARs) in different combinations were successfully produced via regeneration of secondary SEs. The presence of the Chi gene or MARs did not influence the number of transgenic plants produced compared to the marker genes alone. However, the inclusion of MARs contributed to increased mean GFP expression in the population of transgenics. Additionally, the presence of MARs reduced the occurrence of gene silencing and stabilized high levels of GFP expression in lines of transgenic plants multiplied via reiterative somatic embryogenesis. Ninety-four transgenic plants were acclimated in a greenhouse and grown to maturity. Detailed growth analysis indicated that there were no differences in various growth parameters between transgenic and non-transgenic SE-derived plants. Seeds produced from two genetic crosses with one of the transgenic lines were analyzed for EGFP expression-a near-perfect 1:1 segregation was observed, indicating that this line resulted from the insertion of a single locus of T-DNA.

  17. Fast calculation method of computer generated hologram animation for viewpoint parallel shift and rotation using Fourier transform optical system.

    PubMed

    Watanabe, Ryosuke; Yamaguchi, Kazuhiro; Sakamoto, Yuji

    2016-01-20

    Computer generated hologram (CGH) animations can be made by switching many CGHs on an electronic display. Some fast calculation methods for CGH animations have been proposed, but one for viewpoint movement has not been proposed. Therefore, we designed a fast calculation method of CGH animations for viewpoint parallel shifts and rotation. A Fourier transform optical system was adopted to expand the viewing angle. The results of experiments were that the calculation time of our method was over 6 times faster than that of the conventional method. Furthermore, the degradation in CGH animation quality was found to be sufficiently small.

  18. Space-time least-squares finite element method for convection-reaction system with transformed variables

    PubMed Central

    Nam, Jaewook

    2011-01-01

    We present a method to solve a convection-reaction system based on a least-squares finite element method (LSFEM). For steady-state computations, issues related to recirculation flow are stated and demonstrated with a simple example. The method can compute concentration profiles in open flow even when the generation term is small. This is the case for estimating hemolysis in blood. Time-dependent flows are computed with the space-time LSFEM discretization. We observe that the computed hemoglobin concentration can become negative in certain regions of the flow; it is a physically unacceptable result. To prevent this, we propose a quadratic transformation of variables. The transformed governing equation can be solved in a straightforward way by LSFEM with no sign of unphysical behavior. The effect of localized high shear on blood damage is shown in a circular Couette-flow-with-blade configuration, and a physiological condition is tested in an arterial graft flow. PMID:21709752

  19. Review of high-throughput techniques for detecting solid phase Transformation from material libraries produced by combinatorial methods

    NASA Technical Reports Server (NTRS)

    Lee, Jonathan A.

    2005-01-01

    High-throughput measurement techniques are reviewed for solid phase transformation from materials produced by combinatorial methods, which are highly efficient concepts to fabricate large variety of material libraries with different compositional gradients on a single wafer. Combinatorial methods hold high potential for reducing the time and costs associated with the development of new materials, as compared to time-consuming and labor-intensive conventional methods that test large batches of material, one- composition at a time. These high-throughput techniques can be automated to rapidly capture and analyze data, using the entire material library on a single wafer, thereby accelerating the pace of materials discovery and knowledge generation for solid phase transformations. The review covers experimental techniques that are applicable to inorganic materials such as shape memory alloys, graded materials, metal hydrides, ferric materials, semiconductors and industrial alloys.

  20. Development of Marker-Free Insect-Resistant Indica Rice by Agrobacterium tumefaciens-Mediated Co-transformation.

    PubMed

    Ling, Fei; Zhou, Fei; Chen, Hao; Lin, Yongjun

    2016-01-01

    Agrobacterium-mediated co-transformation is an efficient strategy to generate marker-free transgenic plants. In this study, the vectors pMF-2A(∗) containing a synthetic cry2A(∗) gene driven by maize ubiquitin promoter and pCAMBIA1301 harboring hygromycin phosphotransferase gene (hpt) were introduced into Minghui86 (Oryza sativa L. ssp. indica), an elite indica restorer line. Two independent transformants containing both the cry2A(∗) gene and hpt gene were regenerated. Several homozygous marker-free transgenic progenies were derived from family 2AH2, and three of them were selected for further insect bioassay in the laboratory and field. Insect-resistance assays revealed that all the three transgenic lines were highly resistant to striped stem borer (Chilo suppressalis), yellow stem borer (Tryporyza incertulas) and rice leaf folder (Cnaphalocrocis medinalis). The measurement of Cry2A protein concentration showed that Cry2A protein was stably expressed in leaves and stems of homozygous transgenic lines and their hybrids. The yields of the marker-free homozygous transgenic lines and their hybrids were not significantly different from those of their corresponding controls. Furthermore, the results of flanking sequence isolation showed that the T-DNA in line 8-30 was integrated into the intergenic region of chromosome 2 (between Os02g43680 and Os02g43690). These results indicate that the marker-free transgenic rice has the potential for commercial production.

  1. Development of Marker-Free Insect-Resistant Indica Rice by Agrobacterium tumefaciens-Mediated Co-transformation

    PubMed Central

    Ling, Fei; Zhou, Fei; Chen, Hao; Lin, Yongjun

    2016-01-01

    Agrobacterium-mediated co-transformation is an efficient strategy to generate marker-free transgenic plants. In this study, the vectors pMF-2A∗ containing a synthetic cry2A∗ gene driven by maize ubiquitin promoter and pCAMBIA1301 harboring hygromycin phosphotransferase gene (hpt) were introduced into Minghui86 (Oryza sativa L. ssp. indica), an elite indica restorer line. Two independent transformants containing both the cry2A∗ gene and hpt gene were regenerated. Several homozygous marker-free transgenic progenies were derived from family 2AH2, and three of them were selected for further insect bioassay in the laboratory and field. Insect-resistance assays revealed that all the three transgenic lines were highly resistant to striped stem borer (Chilo suppressalis), yellow stem borer (Tryporyza incertulas) and rice leaf folder (Cnaphalocrocis medinalis). The measurement of Cry2A protein concentration showed that Cry2A protein was stably expressed in leaves and stems of homozygous transgenic lines and their hybrids. The yields of the marker-free homozygous transgenic lines and their hybrids were not significantly different from those of their corresponding controls. Furthermore, the results of flanking sequence isolation showed that the T-DNA in line 8-30 was integrated into the intergenic region of chromosome 2 (between Os02g43680 and Os02g43690). These results indicate that the marker-free transgenic rice has the potential for commercial production. PMID:27833629

  2. Transactors, Transformers and Beyond. A Multi-Method Development of a Theoretical Typology of Leadership.

    ERIC Educational Resources Information Center

    Pearce, Craig L.; Sims, Henry P., Jr.; Cox, Jonathan F.; Ball, Gail; Schnell, Eugene; Smith, Ken A.; Trevino, Linda

    2003-01-01

    To extend the transactional-transformational model of leadership, four theoretical behavioral types of leadership were developed based on literature review and data from studies of executive behavior (n=253) and subordinate attitudes (n=208). Confirmatory factor analysis of a third data set (n=702) support the existence of four leadership types:…

  3. Apparatus and method for controlling the temperature of the core of a super-conducting transformer

    SciTech Connect

    Golner, Thomas; Pleva, Edward; Mehta, Shirish

    2006-10-10

    An apparatus for controlling the temperature of a core of a transformer is provided that includes a core, a shield surrounding the core, a cast formed between the core and the shield, and tubing positioned on the shield. The cast directs heat from the core to the shield and cooling fluid is directed through the tubing to cool the shield.

  4. The Federal Transformation Intervention Model in Persistently Lowest Achieving High Schools: A Mixed-Methods Study

    ERIC Educational Resources Information Center

    Le Patner, Michelle B.

    2012-01-01

    This study examined the American Recovery and Reinvestment Act federal mandate of the Transformation Intervention Model (TIM) outlined by the School Improvement Grant, which was designed to turn around persistently lowest achieving schools. The study was conducted in four high schools in a large Southern California urban district that selected the…

  5. Microtiter plate transformation using the LiAc/SS carrier DNA/PEG method.

    PubMed

    Gietz, R Daniel; Schiestl, Robert H

    2007-01-01

    Here, we describe a protocol that has been adapted for the transformation of yeast cells in 96-well microtiter plates. This protocol can be tailored for multiple applications and is suitable for high-throughput applications. It can be completed in 2-3 h, once the yeast cells have been grown depending on the heat shock used.

  6. Setaria viridis floral-dip: A simple and rapid Agrobacterium-medicated transformation method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Setaria viridis was recently described as a new monocotyledonous model species for C4 photosynthesis research and genetic transformation. It has biological attributes (rapid life cycle, small genome, diploid, short stature and simple growth requirements) that make it suitable for use as a model plan...

  7. A High-Throughput Regeneration and Transformation Platform for Production of Genetically Modified Banana.

    PubMed

    Tripathi, Jaindra N; Oduor, Richard O; Tripathi, Leena

    2015-01-01

    Banana (Musa spp.) is an important staple food as well as cash crop in tropical and subtropical countries. Various bacterial, fungal, and viral diseases and pests such as nematodes are major constraints in its production and are currently destabilizing the banana production in sub-Saharan Africa. Genetic engineering is a complementary option used for incorporating useful traits in banana to bypass the long generation time, polyploidy, and sterility of most of the cultivated varieties. A robust transformation protocol for farmer preferred varieties is crucial for banana genomics and improvement. A robust and reproducible system for genetic transformation of banana using embryogenic cell suspensions (ECS) has been developed in this study. Two different types of explants (immature male flowers and multiple buds) were tested for their ability to develop ECS in several varieties of banana locally grown in Africa. ECS of banana varieties "Cavendish Williams" and "Gros Michel" were developed using multiple buds, whereas ECS of "Sukali Ndiizi" was developed using immature male flowers. Regeneration efficiency of ECS was about 20,000-50,000 plantlets per ml of settled cell volume (SCV) depending on variety. ECS of three different varieties were transformed through Agrobacterium-mediated transformation using gusA reporter gene and 20-70 independent transgenic events per ml SCV of ECS were regenerated on selective medium. The presence and integration of gusA gene in transgenic plants was confirmed by PCR, dot blot, and Southern blot analysis and expression by histochemical GUS assays. The robust transformation platform was successfully used to generate hundreds of transgenic lines with disease resistance. Such a platform will facilitate the transfer of technologies to national agricultural research systems (NARS) in Africa.

  8. A High-Throughput Regeneration and Transformation Platform for Production of Genetically Modified Banana

    PubMed Central

    Tripathi, Jaindra N.; Oduor, Richard O.; Tripathi, Leena

    2015-01-01

    Banana (Musa spp.) is an important staple food as well as cash crop in tropical and subtropical countries. Various bacterial, fungal, and viral diseases and pests such as nematodes are major constraints in its production and are currently destabilizing the banana production in sub-Saharan Africa. Genetic engineering is a complementary option used for incorporating useful traits in banana to bypass the long generation time, polyploidy, and sterility of most of the cultivated varieties. A robust transformation protocol for farmer preferred varieties is crucial for banana genomics and improvement. A robust and reproducible system for genetic transformation of banana using embryogenic cell suspensions (ECS) has been developed in this study. Two different types of explants (immature male flowers and multiple buds) were tested for their ability to develop ECS in several varieties of banana locally grown in Africa. ECS of banana varieties “Cavendish Williams” and “Gros Michel” were developed using multiple buds, whereas ECS of “Sukali Ndiizi” was developed using immature male flowers. Regeneration efficiency of ECS was about 20,000–50,000 plantlets per ml of settled cell volume (SCV) depending on variety. ECS of three different varieties were transformed through Agrobacterium-mediated transformation using gusA reporter gene and 20–70 independent transgenic events per ml SCV of ECS were regenerated on selective medium. The presence and integration of gusA gene in transgenic plants was confirmed by PCR, dot blot, and Southern blot analysis and expression by histochemical GUS assays. The robust transformation platform was successfully used to generate hundreds of transgenic lines with disease resistance. Such a platform will facilitate the transfer of technologies to national agricultural research systems (NARS) in Africa. PMID:26635849

  9. A comparison between the method based on a rigorous field analysis and the impedance transformation method for the intrinsic surface impedance of superconductor films

    NASA Astrophysics Data System (ADS)

    Lee, J. H.; Lee, Sang Young

    2006-10-01

    In obtaining the intrinsic surface resistance (RS) from the effective surface resistance (RS,eff) measured at microwave frequencies by using the dielectric resonator method, the impedance transformation method reported by Klein et al. [N. Klein, H. Chaloupka, G. Muller, S. Orbach, H. Piel, B. Roas, L. Schultz, U. Klein, M. Peiniger, J. Appl. Phys. 67 (1990) 6940] has been very useful. Here we compared the RS of YBa2Cu3O7-δ (YBCO) films on dielectric substrates obtained by a rigorous field analysis based on the TE-mode matching method with those by the impedance transformation method. The two methods produced almost the same RS,eff vs. RS relation in most practical cases of the substrate thickness being less than 1 mm and sapphire and rutile used as the materials for the dielectric rod. However, when the resonant frequency of the dielectric resonator became close to that of the resonant structure formed by the substrates and the metallic surroundings, the RS,eff vs. RS relations appeared strikingly different between the two methods. Effects of the TE011-mode cutoff frequency inside the substrate region, which could not be considered in the impedance transformation method, on the relation between the RS,eff and RS of superconductor films are also investigated. We confirmed our arguments by demonstrating a case where existence of evanescent modes should be considered for obtaining the RS of YBCO films from the RS,eff.

  10. Remote sensing image fusion method in CIELab color space using nonsubsampled shearlet transform and pulse coupled neural networks

    NASA Astrophysics Data System (ADS)

    Jin, Xin; Zhou, Dongming; Yao, Shaowen; Nie, Rencan; Yu, Chuanbo; Ding, Tingting

    2016-04-01

    In CIELab color space, we propose a remote sensing image fusion technique based on nonsubsampled shearlet transform (NSST) and pulse coupled neural network (PCNN), which aim to improve the efficiency and performance of the remote sensing image fusion by combining the excellent properties of the two methods. First, panchromatic (PAN) and multispectral (MS) are transformed into CIELab color space to get different color components. Second, PAN and L component of MS are decomposed by the NSST to obtain corresponding the low-frequency coefficients and high-frequency coefficients. Third, the low-frequency coefficients are fused by intersecting cortical model (ICM); the high-frequency coefficients are divided into several sub-blocks to calculate the average gradient (AG), and the linking strength β of PCNN model is determined by the AG, so that the parameters β can be adaptively set according to the quality of the sub-block images, then the sub-blocks image are input into PCNN to get the oscillation frequency graph (OFG), the method can get the fused high-frequency coefficients according to the OFG. Finally, the fused L component is obtained by inverse NSST, and the fused RGB color image is obtained through inverse CIELab transform. The experimental results demonstrate that the proposed method provide better effect compared with other common methods.

  11. A Suitable Coordinate Transformation Method for Correcting Voltage Vector in Motor Current Detection Using a Single Shunt Resistor

    NASA Astrophysics Data System (ADS)

    Tomigashi, Yoshio; Hida, Hajime; Ueyama, Kenji

    To reduce costs of inverters, a current detection method using a single shunt resistor is required for motor drive systems in home electrical appliances. In this paper, a method is proposed to correct a voltage reference vector by converting coordinates from a rotating reference frame into a fixed reference frame. Also proposed is a new coordinate transformation method that is appropriate for the correction. Authors focused on the undetectable area that exists every 60 degrees in α-β coordinates. When the α-β coordinates in an nπ/3 rotation are defined as αn-βn, the αn-axis can be defined as the central axis in an undetectable area. We propose a coordinate transformation method that converts the voltage vector in the d-q coordinates into uvw phase voltages through αn-βn coordinates then correct it. This method corrects the voltage vector by a very simple algorithm that limits the αn-βn elements. The effectiveness of the proposed method is confirmed by simulation and experiment. Currents were clearly detected by using the proposed method. This shows that the proposed method is suitable for position sensor-less drives in permanent magnet synchronous motors.

  12. Entropy generation in Gaussian quantum transformations: applying the replica method to continuous-variable quantum information theory

    NASA Astrophysics Data System (ADS)

    Gagatsos, Christos N.; Karanikas, Alexandros I.; Kordas, Georgios; Cerf, Nicolas J.

    2016-02-01

    In spite of their simple description in terms of rotations or symplectic transformations in phase space, quadratic Hamiltonians such as those modelling the most common Gaussian operations on bosonic modes remain poorly understood in terms of entropy production. For instance, determining the quantum entropy generated by a Bogoliubov transformation is notably a hard problem, with generally no known analytical solution, while it is vital to the characterisation of quantum communication via bosonic channels. Here we overcome this difficulty by adapting the replica method, a tool borrowed from statistical physics and quantum field theory. We exhibit a first application of this method to continuous-variable quantum information theory, where it enables accessing entropies in an optical parametric amplifier. As an illustration, we determine the entropy generated by amplifying a binary superposition of the vacuum and a Fock state, which yields a surprisingly simple, yet unknown analytical expression.

  13. Analytical Solutions for the Resonance Response of Goupillaud-type Elastic Media Using Z-transform Methods

    DTIC Science & Technology

    2012-02-01

    public release; distribution is unlimited. NOTICES Disclaimers The findings in this report are not to be construed as an official Department of...using z-transform methods. The determinant of the resulting global system matrix in the z-space |Am| is a palindromic polynomial with real...Alcalá Park, San Diego, CA 92110, USA a r t i c l e i n f o Article history: Received 14 March 2011 Received in revised form 1 August 2011 Accepted 5

  14. [Construction of a bivalent plant expression vector carrying VvSUC11 and VvSUC12 genes and its genetic transformation in sugar beet].

    PubMed

    Yin, Donglin; Zhu, Jianbo; Wang, Aiying; Xiang, Benchun

    2011-08-01

    We have recombined genes VvSUC11, VvSUC12 from Vitis vinifera L., and root-specific promoters of sweet potato storage protein gene from Ipomoea batatas L. Lam., named as SP1 and SP2. We have constructed a vector pCAMBIA2301-SP1- VvSUC11-SP2-VvSUC12 using pCAMBIA2301 as an original vector. VvSUC11 and VvSUC12 were under the control of root-specific promoters of sweet potato storage protein gene. We transformed the vector into KWS-9103 breeding line of Beta vulgaris L. with Agrobacterium-mediated transformation. We have established the optimal genetic transformation protocol of sugar beet as following: the explants pre-cultured for 4 days were immersed in Agrobacterium suspension of OD(600)=0.5, supplemented with 0.005% Silwet L-77, and followed by a 4-day culture on medium containing cefotaxime, then the buds were selected on medium containing kanamycin and cefotaxime. The percentage of kanamycin-resistant buds was as high as 42%. Results of PCR and RT-PCR proved that the target genes had integrated into sugar beet genome and expressed. It will lay a foundation for further studying their function in Beta vulgaris.

  15. Co-transforming bar and CsLEA enhanced tolerance to drought and salt stress in transgenic alfalfa (Medicago sativa L.).

    PubMed

    Zhang, Jiyu; Duan, Zhen; Zhang, Daiyu; Zhang, Jianquan; Di, Hongyan; Wu, Fan; Wang, Yanrong

    2016-03-25

    Drought and high salinity are two major abiotic factors that restrict alfalfa productivity. A dehydrin protein, CsLEA, from the desert grass Cleistogenes songorica was transformed into alfalfa (Medicago sativa L.) via Agrobacterium-mediated transformation using the bar gene as a selectable marker, and the drought and salt stress tolerances of the transgenic plants were assessed. Thirty-nine of 119 transformants were positive, as screened by Basta, and further molecularly authenticated using PCR and RT-PCR. Phenotype observations revealed that the transgenic plants grew better than the wild-type (WT) plants after 15d of drought stress and 10d of salt stress: the leaves of WT alfalfa turned yellow, whereas the transgenic alfalfa leaves only wilted; after rewatering, the transgenic plants returned to a normal state, though the WT plants could not be restored. Evaluation of physiologic and biochemical indices during drought and salt stresses showed a relatively lower Na(+) content in the leaves of the transgenic plants, which would reduce toxic ion effects. In addition, the transgenic plants were able to maintain a higher relative water content (RWC), higher shoot biomass, fewer photosystem changes, decreased membrane injury, and a lower level of osmotic stress injury. These results demonstrate that overexpression of the CsLEA gene can enhance the drought and salt tolerance of transgenic alfalfa; in addition, carrying the bar gene in the genome may increase herbicide resistance.

  16. A genetic-algorithm-based method to find unitary transformations for any desired quantum computation and application to a one-bit oracle decision problem

    NASA Astrophysics Data System (ADS)

    Bang, Jeongho; Yoo, Seokwon

    2014-12-01

    We propose a genetic-algorithm-based method to find the unitary transformations for any desired quantum computation. We formulate a simple genetic algorithm by introducing the "genetic parameter vector" of the unitary transformations to be found. In the genetic algorithm process, all components of the genetic parameter vectors are supposed to evolve to the solution parameters of the unitary transformations. We apply our method to find the optimal unitary transformations and to generalize the corresponding quantum algorithms for a realistic problem, the one-bit oracle decision problem, or the often-called Deutsch problem. By numerical simulations, we can faithfully find the appropriate unitary transformations to solve the problem by using our method. We analyze the quantum algorithms identified by the found unitary transformations and generalize the variant models of the original Deutsch's algorithm.

  17. A new multi-step technique with differential transform method for analytical solution of some nonlinear variable delay differential equations.

    PubMed

    Benhammouda, Brahim; Vazquez-Leal, Hector

    2016-01-01

    This work presents an analytical solution of some nonlinear delay differential equations (DDEs) with variable delays. Such DDEs are difficult to treat numerically and cannot be solved by existing general purpose codes. A new method of steps combined with the differential transform method (DTM) is proposed as a powerful tool to solve these DDEs. This method reduces the DDEs to ordinary differential equations that are then solved by the DTM. Furthermore, we show that the solutions can be improved by Laplace-Padé resummation method. Two examples are presented to show the efficiency of the proposed technique. The main advantage of this technique is that it possesses a simple procedure based on a few straight forward steps and can be combined with any analytical method, other than the DTM, like the homotopy perturbation method.

  18. Agrobacterium tumefaciens-mediated sorghum transformation using a mannose selection system.

    PubMed

    Gao, Zhensheng; Xie, Xueju; Ling, Yan; Muthukrishnan, Subbarat; Liang, George H

    2005-11-01

    A dual-marker plasmid containing the selectable marker gene, manA, and the reporter gene, sgfp, was used to transform immature sorghum embryos by employing an Agrobacterium-mediated system. Both genes were under the control of the ubi1 promoter in a binary vector pPZP201. The Escherichia coli phosphomannose isomerase (PMI) gene, pmi, was used as the selectable marker gene and mannose was used as the selective agent. The sgfp gene encoding green fluorescence protein (GFP) was the reporter gene and served as a visual screening marker. A total of 167 transgenic plants were obtained from nine different embryogenic callus lines grown on a selection medium containing 1%-2% mannose. Embryoids and shoots regenerated via embryogenesis, that showed strong GFP fluorescence, were selected from two sorghum genotypes: C401, an inbred line, and Pioneer 8505, a commercial hybrid. The GFP accumulation in transgenic plants was observed with a dissecting stereomicroscope. The integration and expression of the manA gene was confirmed by Southern blot and Western blot analyses, and the feasibility of manA selection was demonstrated by the chlorophenol red (CPR) assay. Our results indicated that transgenes segregated in the Mendelian fashion in the T1 generation. The conversion of mannose to a metabolizable fructose carbon source is beneficial to plants. In addition, except in soybean and a few legumes, no endogenous PMI activity has been detected in plant species, indicating that PMI is useful in the transformation of sorghum. In addition, PMI has no sequence homology to known allergens. Optimization of this selection system for sorghum transformation provides an efficient way to produce transgenic plants without using antibiotic or herbicidal agents as selectable markers, and our results showed that the transformation efficiency reached 2.88% for Pioneer 8505 and 3.30% for C401, both values higher than in previously published reports.

  19. Tensor Product Model Transformation Based Adaptive Integral-Sliding Mode Controller: Equivalent Control Method

    PubMed Central

    Zhao, Guoliang; Li, Hongxing

    2013-01-01

    This paper proposes new methodologies for the design of adaptive integral-sliding mode control. A tensor product model transformation based adaptive integral-sliding mode control law with respect to uncertainties and perturbations is studied, while upper bounds on the perturbations and uncertainties are assumed to be unknown. The advantage of proposed controllers consists in having a dynamical adaptive control gain to establish a sliding mode right at the beginning of the process. Gain dynamics ensure a reasonable adaptive gain with respect to the uncertainties. Finally, efficacy of the proposed controller is verified by simulations on an uncertain nonlinear system model. PMID:24453897

  20. Tensor product model transformation based adaptive integral-sliding mode controller: equivalent control method.

    PubMed

    Zhao, Guoliang; Sun, Kaibiao; Li, Hongxing

    2013-01-01

    This paper proposes new methodologies for the design of adaptive integral-sliding mode control. A tensor product model transformation based adaptive integral-sliding mode control law with respect to uncertainties and perturbations is studied, while upper bounds on the perturbations and uncertainties are assumed to be unknown. The advantage of proposed controllers consists in having a dynamical adaptive control gain to establish a sliding mode right at the beginning of the process. Gain dynamics ensure a reasonable adaptive gain with respect to the uncertainties. Finally, efficacy of the proposed controller is verified by simulations on an uncertain nonlinear system model.

  1. Estimation of Instantaneous Gas Exchange in Flow-Through Respirometry Systems: A Modern Revision of Bartholomew's Z-Transform Method.

    PubMed

    Pendar, Hodjat; Socha, John J

    2015-01-01

    Flow-through respirometry systems provide accurate measurement of gas exchange over long periods of time. However, these systems have limitations in tracking rapid changes. When an animal infuses a metabolic gas into the respirometry chamber in a short burst, diffusion and airflow in the chamber gradually alter the original signal before it arrives at the gas analyzer. For single or multiple bursts, the recorded signal is smeared or mixed, which may result in dramatically altered recordings compared to the emitted signal. Recovering the original metabolic signal is a difficult task because of the inherent ill conditioning problem. Here, we present two new methods to recover the fast dynamics of metabolic patterns from recorded data. We first re-derive the equations of the well-known Z-transform method (ZT method) to show the source of imprecision in this method. Then, we develop a new model of analysis for respirometry systems based on the experimentally determined impulse response, which is the response of the system to a very short unit input. As a result, we present a major modification of the ZT method (dubbed the 'EZT method') by using a new model for the impulse response, enhancing its precision to recover the true metabolic signals. The second method, the generalized Z-transform (GZT) method, was then developed by generalizing the EZT method; it can be applied to any flow-through respirometry system with any arbitrary impulse response. Experiments verified that the accuracy of recovering the true metabolic signals is significantly improved by the new methods. These new methods can be used more broadly for input estimation in variety of physiological systems.

  2. A quantitative method to detect explosives and selected semivolatiles in soil samples by Fourier transform infrared spectroscopy

    SciTech Connect

    Clapper-Gowdy, M.; Dermirgian, J.; Robitaille, G.

    1995-06-01

    This paper describes a novel Fourier transform infrared (FTIR) spectroscopic method that can be used to rapidly screen soil samples from potentially hazardous waste sites. Samples are heated in a thermal desorption unit and the resultant vapors are collected and analyzed in a long-path gas cell mounted in a FTIR. Laboratory analysis of a soil sample by FTIR takes approximately 10 minutes. This method has been developed to identify and quantify microgram concentrations of explosives in soil samples and is directly applicable to the detection of selected volatile organics, semivolatile organics, and pesticides.

  3. Use of the generalized integral transform method for solving equations of solute transport in porous media

    NASA Astrophysics Data System (ADS)

    Liu, Chongxuan; Szecsody, Jim E.; Zachara, John M.; Ball, William P.

    The generalized integral transform technique (GITT) is applied to solve the one-dimensional advection-dispersion equation (ADE) in heterogeneous porous media coupled with either linear or nonlinear sorption and decay. When both sorption and decay are linear, analytical solutions are obtained using the GITT for one-dimensional ADEs with spatially and temporally variable flow and dispersion coefficient and arbitrary initial and boundary conditions. When either sorption or decay is nonlinear the solutions to ADEs with the GITT are hybrid analytical-numerical. In both linear and nonlinear cases, the forward and inverse integral transforms for the problems described in the paper are apparent and straightforward. Some illustrative examples with linear sorption and decay are presented to demonstrate the application and check the accuracy of the derived analytical solutions. The derived hybrid analytical-numerical solutions are checked against a numerical approach and demonstratively applied to a nonlinear transport example, which simulates a simplified system of iron oxide bioreduction with nonlinear sorption and nonlinear reaction kinetics.

  4. Transforming incomplete fault tree to Ishikawa diagram as an alternative method for technology transfer

    NASA Astrophysics Data System (ADS)

    Batzias, Dimitris F.

    2012-12-01

    Fault Tree Analysis (FTA) can be used for technology transfer when the relevant problem (called 'top even' in FTA) is solved in a technology centre and the results are diffused to interested parties (usually Small Medium Enterprises - SMEs) that have not the proper equipment and the required know-how to solve the problem by their own. Nevertheless, there is a significant drawback in this procedure: the information usually provided by the SMEs to the technology centre, about production conditions and corresponding quality characteristics of the product, and (sometimes) the relevant expertise in the Knowledge Base of this centre may be inadequate to form a complete fault tree. Since such cases are quite frequent in practice, we have developed a methodology for transforming incomplete fault tree to Ishikawa diagram, which is more flexible and less strict in establishing causal chains, because it uses a surface phenomenological level with a limited number of categories of faults. On the other hand, such an Ishikawa diagram can be extended to simulate a fault tree as relevant knowledge increases. An implementation of this transformation, referring to anodization of aluminium, is presented.

  5. Study of {sup 3}He(e,e{prime}) longitudinal response functions with the integral-transform method

    SciTech Connect

    Dobretsov, V.Yu.; Efros, V.D.; Shao, B.

    1995-09-01

    The method of integral transforms is first applied to study of the {sup 3}He longitudinal response functions. The transforms are calculated from localized bound-state type solutions to an inhomogenous three-body equation. {sup 3}N dynamics and the conventional charge density operator serve as an input. The final-state interaction the contribution of the T = 3/2 final states to the problem suppressed and amounts to about 15%. This might be ascribed to symmetry properties of the final-state wave functions. The contributions of the p-wave N/N interaction and of the Coulomb interaction to the problem are studied and found to amount to several percent. Uncertainties due to different choices of s-wave NN forces are found to be of a similar magnitude. For q = 300 MeV/c, the results obtained agree with experiment. For q = 500 MeV/c, noticeable differences are detected. 20 refs., 3 figs.

  6. Estimation of Instantaneous Gas Exchange in Flow-Through Respirometry Systems: A Modern Revision of Bartholomew's Z-Transform Method

    PubMed Central

    Pendar, Hodjat; Socha, John J.

    2015-01-01

    Flow-through respirometry systems provide accurate measurement of gas exchange over long periods of time. However, these systems have limitations in tracking rapid changes. When an animal infuses a metabolic gas into the respirometry chamber in a short burst, diffusion and airflow in the chamber gradually alter the original signal before it arrives at the gas analyzer. For single or multiple bursts, the recorded signal is smeared or mixed, which may result in dramatically altered recordings compared to the emitted signal. Recovering the original metabolic signal is a difficult task because of the inherent ill conditioning problem. Here, we present two new methods to recover the fast dynamics of metabolic patterns from recorded data. We first re-derive the equations of the well-known Z-transform method (ZT method) to show the source of imprecision in this method. Then, we develop a new model of analysis for respirometry systems based on the experimentally determined impulse response, which is the response of the system to a very short unit input. As a result, we present a major modification of the ZT method (dubbed the ‘EZT method’) by using a new model for the impulse response, enhancing its precision to recover the true metabolic signals. The second method, the generalized Z-transform (GZT) method, was then developed by generalizing the EZT method; it can be applied to any flow-through respirometry system with any arbitrary impulse response. Experiments verified that the accuracy of recovering the true metabolic signals is significantly improved by the new methods. These new methods can be used more broadly for input estimation in variety of physiological systems. PMID:26466361

  7. A Restoration of the Image Contaminated with the Photon Shot Noise by an M-Transform Method

    NASA Astrophysics Data System (ADS)

    Hoshino, Kazuhiro; Takaichi, Masato; Nishimura, Toshihiro

    We propose a novel approach for the photon shot noise reduction in image sensors inspired from an M-transform method. In our proposed method, the photon shot noise is transfered to small amplitude random signals. Then the small amplitude random signals are removed by the ε filter. The Sobel method was used for the edge detection and the edge was preserved by not applying the ε filter into the edge. After the noise reduction, the signals are restored to the image by a reverse M-transform. We compared our method with the smoothing filter, the median filter and the Wiener filter. In the photon shot noise according to the Poisson distribution, we changed the incidence number of photons and created the noise image by the simulation. In cases where the number of the maximum incidence photons was 128 pieces (illumination of about 2 lux), the proposal technique indicated the PSNR (Peak-Signal to Noise Ratio) value better than the Wiener filter. Furthermore, the proposal technique confirmed better than the smoothing filter subjectively according to the effect of edge preservation also under dark environment. As the result, our method is confirmed effective for the photon shot noise reduction.

  8. Transport velocity transformation - A convenient method for performance analysis of multilayer solar cell structure

    NASA Technical Reports Server (NTRS)

    Wolf, M.

    1981-01-01

    It is noted that in the case of low-level injection, space-charge quasi-neutrality, and spatially constant material parameters (including an electrostatic field), the individual layer can be treated analytically and the basic solar cell performance parameters can be evaluated from three equations. The first equation represents the transformation of the transport velocity across the layer from the other layer boundary. The second establishes the light-generated current output from the layer interface, under the influence of the transport velocities and minority-carrier density at both layer boundaries and of bulk recombination. The third equation describes the flow of these carriers across other layers. The power of the approach is considered to lie in its facility for analysis of the solar cell's performance layer by layer, giving a clear picture of the individual layer's influence on cell efficiency.

  9. Transform methods for developing parallel algorithms for cyclic-block signal processing

    NASA Astrophysics Data System (ADS)

    Marshall, T. G., Jr.

    A class of FIR and IIR single and multirate parallel filtering algorithms is introduced in which blocks of inputs and outputs are processed on-the-fly in a cyclic manner. There is no inherent latency introduced by the decomposition procedure giving the parallelism, the system latency being primarily due to the component processors. The structure is particularly well-suited for systems in which the component processors are the familiar DSP chips optimized for convolution although other component structures can be accommodated. In particular, the automatic data shifting feature of the TMS320 series processors can be utilized in these algorithms. A transform notation, introduced for digital filter banks, is recast in the desired form for this application. The resulting structure of the system, in this notation, is a circulant matrix for FIR filtering or a related matrix in other cases. The cyclic properties of the system and useful implementation flexibility result from this matrix structure.

  10. Effect of Dispersion Method on Stability and Dielectric Strength of Transformer Oil-Based TiO2 Nanofluids.

    PubMed

    Lv, Yu-Zhen; Li, Chao; Sun, Qian; Huang, Meng; Li, Cheng-Rong; Qi, Bo

    2016-12-01

    Dispersion stability of nanoparticles in the liquid media is of great importance to the utilization in practice. This study aims to investigate the effects of mechanical dispersion method on the dispersibility of functionalized TiO2 nanoparticles in the transformer oil. Dispersion methods, including stirring, ultrasonic bath, and probe processes, were systematically tested to verify their versatility for preparing stable nanofluid. The test results reveal that the combination of ultrasonic bath process and stirring method has the best dispersion efficiency and the obtained nanofluid possesses the highest AC breakdown strength. Specifically, after aging for 168 h, the size of nanoparticles in the nanofluid prepared by the combination method has no obvious change, while those obtained by the other three paths are increased obviously.

  11. Effect of Dispersion Method on Stability and Dielectric Strength of Transformer Oil-Based TiO2 Nanofluids

    NASA Astrophysics Data System (ADS)

    Lv, Yu-zhen; Li, Chao; Sun, Qian; Huang, Meng; Li, Cheng-rong; Qi, Bo

    2016-11-01

    Dispersion stability of nanoparticles in the liquid media is of great importance to the utilization in practice. This study aims to investigate the effects of mechanical dispersion method on the dispersibility of functionalized TiO2 nanoparticles in the transformer oil. Dispersion methods, including stirring, ultrasonic bath, and probe processes, were systematically tested to verify their versatility for preparing stable nanofluid. The test results reveal that the combination of ultrasonic bath process and stirring method has the best dispersion efficiency and the obtained nanofluid possesses the highest AC breakdown strength. Specifically, after aging for 168 h, the size of nanoparticles in the nanofluid prepared by the combination method has no obvious change, while those obtained by the other three paths are increased obviously.

  12. A combination of dual-tree discrete wavelet transform and minimum redundancy maximum relevance method for diagnosis of Alzheimer's disease.

    PubMed

    Aggarwal, Namita; Rana, Bharti; Agrawal, R K; Kumaran, Senthil

    2015-01-01

    In this paper, we propose a three-phased method for diagnosis of Alzheimer's disease using the structural magnetic resonance imaging (MRI). In first phase, gray matter tissue probability map is obtained from every brain MRI volume. Further, five regions of interest (ROIs) are extracted as per prior knowledge. In second phase, features are extracted from each ROI using 3D dual-tree discrete wavelet transform. In third phase, relevant features are selected using minimum redundancy maximum relevance features selection technique. The decision model is built with features so obtained, using a classifier. To evaluate the effectiveness of the proposed method, experiments are performed with four well-known classifiers on four data sets, built from a publicly available OASIS database. The performance is evaluated in terms of sensitivity, specificity and classification accuracy. It was observed that the proposed method outperforms existing methods in terms of all three performance measures. This is further validated with statistical tests.

  13. Analytical energy gradient based on spin-free infinite-order Douglas-Kroll-Hess method with local unitary transformation.

    PubMed

    Nakajima, Yuya; Seino, Junji; Nakai, Hiromi

    2013-12-28

    In this study, the analytical energy gradient for the spin-free infinite-order Douglas-Kroll-Hess (IODKH) method at the levels of the Hartree-Fock (HF), density functional theory (DFT), and second-order Møller-Plesset perturbation theory (MP2) is developed. Furthermore, adopting the local unitary transformation (LUT) scheme for the IODKH method improves the efficiency in computation of the analytical energy gradient. Numerical assessments of the present gradient method are performed at the HF, DFT, and MP2 levels for the IODKH with and without the LUT scheme. The accuracies are examined for diatomic molecules such as hydrogen halides, halogen dimers, coinage metal (Cu, Ag, and Au) halides, and coinage metal dimers, and 20 metal complexes, including the fourth-sixth row transition metals. In addition, the efficiencies are investigated for one-, two-, and three-dimensional silver clusters. The numerical results confirm the accuracy and efficiency of the present method.

  14. A method for detection and characterisation of structural non-linearities using the Hilbert transform and neural networks

    NASA Astrophysics Data System (ADS)

    Ondra, V.; Sever, I. A.; Schwingshackl, C. W.

    2017-01-01

    This paper presents a method for detection and characterisation of structural non-linearities from a single frequency response function using the Hilbert transform in the frequency domain and artificial neural networks. A frequency response function is described based on its Hilbert transform using several common and newly introduced scalar parameters, termed non-linearity indexes, to create training data of the artificial neural network. This network is subsequently used to detect the existence of non-linearity and classify its type. The theoretical background of the method is given and its usage is demonstrated on different numerical test cases created by single degree of freedom non-linear systems and a lumped parameter multi degree of freedom system with a geometric non-linearity. The method is also applied to several experimentally measured frequency response functions obtained from a cantilever beam with a clearance non-linearity and an under-platform damper experimental rig with a complex friction contact interface. It is shown that the method is a fast and noise-robust means of detecting and characterising non-linear behaviour from a single frequency response function.

  15. [Determination of partition coefficient of dissolved gases in transformer oil using phase ratio variation method and static headspace gas chromatography].

    PubMed

    Zhao, Jinghong; Wang, Hailong; Liu, Wenmin; Zhou, Yansheng; Guan, Yafeng

    2004-05-01

    The partition coefficients of dissolved gases in transformer oil were determined using a phase ratio variation method and static headspace gas chromatography (GC). A pressure balancing and gas volume-metering device was connected to the vent of a sample loop on a six-port injection valve of the GC. The gas phase sample from the headspace vial of 25 mL was transferred to an 80 microL sample-loop through a fused silica capillary of 0.53 mm i.d., and then separated and determined quantitatively by GC. A 2 m x 1 mm i.d. GDX502 micro-packed column was used for the separation. Five different gas-liquid volume ratios in the headspace vials were measured at different equilibrium concentrations. The partition coefficients of hydrocarbon gases including methane, acetylene, ethylene, ethane and propane dissolved in transformer oil were determined by using linear regression analysis at 20 degrees C and 50 degrees C separately. The errors between the real values and regression values from experimental data were less than 4.14% except methane. Fundamental data for on-line measurement of dissolved gases in transformer oil are provided by GC.

  16. Transformational Learners: Transformational Teachers

    ERIC Educational Resources Information Center

    Jones, Marguerite

    2009-01-01

    Transformational learning, according to Mezirow (1981), involves transforming taken-for-granted frames of reference into more discriminating, flexible "habits of mind". In teacher education, transformative learning impacts on the development of students' action theories, self-efficacy and professional attributes. Although considered…

  17. Agrobacterium-transformed rice plants expressing synthetic cryIA(b) and cryIA(c) genes are highly toxic to striped stem borer and yellow stem borer.

    PubMed

    Cheng, X; Sardana, R; Kaplan, H; Altosaar, I

    1998-03-17

    Over 2,600 transgenic rice plants in nine strains were regenerated from >500 independently selected hygromycin-resistant calli after Agrobacterium-mediated transformation. The plants were transformed with fully modified (plant codon optimized) versions of two synthetic cryIA(b) and cryIA(c) coding sequences from Bacillus thuringiensis as well as the hph and gus genes, coding for hygromycin phosphotransferase and beta-glucuronidase, respectively. These sequences were placed under control of the maize ubiquitin promoter, the CaMV35S promoter, and the Brassica Bp10 gene promoter to achieve high and tissue-specific expression of the lepidopteran-specific delta-endotoxins. The integration, expression, and inheritance of these genes were demonstrated in R0 and R1 generations by Southern, Northern, and Western analyses and by other techniques. Accumulation of high levels (up to 3% of soluble proteins) of CryIA(b) and CryIA(c) proteins was detected in R0 plants. Bioassays with R1 transgenic plants indicated that the transgenic plants were highly toxic to two major rice insect pests, striped stem borer (Chilo suppressalis) and yellow stem borer (Scirpophaga incertulas), with mortalities of 97-100% within 5 days after infestation, thus offering a potential for effective insect resistance in transgenic rice plants.

  18. Method for transforming CT images for attenuation correction in PET/CT imaging

    SciTech Connect

    Carney, Jonathan P.J.; Townsend, David W.; Rappoport, Vitaliy; Bendriem, Bernard

    2006-04-15

    A tube-voltage-dependent scheme is presented for transforming Hounsfield units (HU) measured by different computed tomography (CT) scanners at different x-ray tube voltages (kVp) to 511 keV linear attenuation values for attenuation correction in positron emission tomography (PET) data reconstruction. A Gammex 467 electron density CT phantom was imaged using a Siemens Sensation 16-slice CT, a Siemens Emotion 6-slice CT, a GE Lightspeed 16-slice CT, a Hitachi CXR 4-slice CT, and a Toshiba Aquilion 16-slice CT at kVp ranging from 80 to 140 kVp. All of these CT scanners are also available in combination with a PET scanner as a PET/CT tomograph. HU obtained for various reference tissue substitutes in the phantom were compared with the known linear attenuation values at 511 keV. The transformation, appropriate for lung, soft tissue, and bone, yields the function 9.6x10{sup -5}{center_dot}(HU+1000) below a threshold of {approx}50 HU and a{center_dot}(HU+1000)+b above the threshold, where a and b are fixed parameters that depend on the kVp setting. The use of the kVp-dependent scaling procedure leads to a significant improvement in reconstructed PET activity levels in phantom measurements, resolving errors of almost 40% otherwise seen for the case of dense bone phantoms at 80 kVp. Results are also presented for patient studies involving multiple CT scans at different kVp settings, which should all lead to the same 511 keV linear attenuation values. A linear fit to values obtained from 140 kVp CT images using the kVp-dependent scaling plotted as a function of the corresponding values obtained from 80 kVp CT images yielded y=1.003x-0.001 with an R{sup 2} value of 0.999, indicating that the same values are obtained to a high degree of accuracy.

  19. A Bridge Health Monitoring Method Using the Hilbert-Huang Transform: A Case Study

    NASA Technical Reports Server (NTRS)

    Huang, Norden E.; Chiang, Weiling; Busalacchi, Antonio J. (Technical Monitor)

    2000-01-01

    We have developed a new method for nondestructive instrument method to monitor the health of a bridge. This new method is based on a transient test load and simple data collection. The nuclear of the method is the new invented nonstationary and nonlinear time series analysis method, the Empirical Mode Decomposition and Hilbert Spectral Analysis. The final decision on the health of the bridge structure is based on the nonlinear characteristic of the data, and on the comparison between the free and the forced vibration frequencies. Thus this alternative method enjoys many advantages: (1) no a priori data required, (2) simple data collection, and (3) minimum traffic disruption. Results from a case study of the Shing-Nan Bridge in Hou-Wei will be reported.

  20. Transforming Self and Others through Research: Transpersonal Research Methods and Skills for the Human Sciences and Humanities. SUNY Series in Transpersonal and Humanistic Psychology

    ERIC Educational Resources Information Center

    Anderson, Rosemarie; Braud, William

    2011-01-01

    Research approaches in the field of transpersonal psychology can be transformative for researchers, participants, and the audience of a project. This book offers these transformative approaches to those conducting research across the human sciences and the humanities. Rosemarie Anderson and William Braud first described such methods in…

  1. Density of band-gap traps in polycrystalline films from photoconductivity transients using an improved Laplace transform method

    NASA Astrophysics Data System (ADS)

    Studenikin, S. A.; Golego, Nickolay; Cocivera, Michael

    1998-11-01

    Slow relaxation of the photoconductivity over a period of days was studied in polycrystalline ZnO and TiO2 films prepared by spray pyrolysis. The phenomenon is described by a model involving deep sensitizing hole traps in the forbidden gap. The trap state distribution based on this model was calculated using an improved Laplace transform method. For ZnO and TiO2 films, the density of states was shown to have a peak-like distribution with a maximum near the lower third of the energy gap.

  2. Pretreatment method for immunoassay of polychlorinated biphenyls in transformer oil using multilayer capillary column and microfluidic liquid-liquid partitioning.

    PubMed

    Aota, Arata; Date, Yasumoto; Terakado, Shingo; Ohmura, Naoya

    2013-01-01

    Polychlorinated biphenyls (PCBs) are persistent organic pollutants that are present in the insulating oil inside a large number of transformers. To aid in eliminating PCB-contaminated transformers, PCBs in oil need to be measured using a rapid and cost-effective analytical method. We previously reported a pretreatment method for the immunoassay of PCBs in oil using a large-scale multilayer column and a microchip with multiple microrecesses, which permitted concentrated solvent extraction. In this paper, we report on a more rapid and facile pretreatment method, without an evaporation process, by improving the column and the microchip. In a miniaturized column, the decomposition and separation of oil were completed in 2 min. PCBs can be eluted from the capillary column at concentrations seven-times higher than those from the previous column. The total volume of the microrecesses was increased by improving the microrecess structure, the enabling extraction of four-times the amount of PCBs achieved with the previous system. By interfacing the capillary column with the improved microchip, PCBs in the eluate from the column were extracted into dimethyl sulfoxide in microrecesses with high enrichment and without the need for evaporation. Pretreatment was completed within 20 min. The pretreated oil was analyzed using a flow-based kinetic exclusion immunoassay. The limit of detection of PCBs in oil was 0.15 mg kg(-1), which satisfies the criterion set in Japan of 0.5 mg kg(-1).

  3. Wavelet transform-based methods for denoising of Coulter counter signals

    NASA Astrophysics Data System (ADS)

    Jagtiani, Ashish V.; Sawant, Rupesh; Carletta, Joan; Zhe, Jiang

    2008-06-01

    A process based on discrete wavelet transforms is developed for denoising and baseline correction of measured signals from Coulter counters. Given signals from a particular Coulter counting experiment, which detect passage of particles through a fluid-filled microchannel, the process uses a cross-validation procedure to pick appropriate parameters for signal denoising; these parameters include the choice of the particular wavelet, the number of levels of decomposition, the threshold value and the threshold strategy. The process is demonstrated on simulated and experimental single channel data obtained from a particular multi-channel Coulter counter processing. For these example experimental signals from 20 µm polymethacrylate and Cottonwood/Eastern Deltoid pollen particles and the simulated signals, denoising is aimed at removing Gaussian white noise, 60 Hz power line interference and low frequency baseline drift. The process can be easily adapted for other Coulter counters and other sources of noise. Overall, wavelets are presented as a tool to aid in accurate detection of particles in Coulter counters.

  4. The method of unitary clothing transformations in the theory of nucleon-nucleon scattering

    NASA Astrophysics Data System (ADS)

    Dubovyk, I.; Shebeko, A.

    2010-04-01

    The clothing procedure, put forward in quantum field theory (QFT) by Greenberg and Schweber, is applied for the description of nucleon-nucleon (N -N) scattering. We consider pseudoscalar (π and η), vector (ρ and ω) and scalar (δ and σ) meson fields interacting with 1/2 spin (N and N) fermion ones via the Yukawa-type couplings to introduce trial interactions between “bare” particles. The subsequent unitary clothing transformations (UCTs) are found to express the total Hamiltonian through new interaction operators that refer to particles with physical (observable) properties, the so-called clothed particles. In this work, we are focused upon the Hermitian and energy-independent operators for the clothed nucleons, being built up in the second order in the coupling constants. The corresponding analytic expressions in momentum space are compared with the separate meson contributions to the one-boson-exchange potentials in the meson theory of nuclear forces. In order to evaluate the T matrix of the N-N scattering we have used an equivalence theorem that enables us to operate in the clothed particle representation (CPR) instead of the bare particle representation (BPR) with its huge amount of virtual processes. We have derived the Lippmann-Schwinger(LS)-type equation for the CPR elements of the T-matrix for a given collision energy in the two-nucleon sector of the Hilbert space H of hadronic states and elaborated a code for its numerical solution in momentum space.

  5. A simple method to fabricate lenses for in situ Fourier transform infrared spectroscopy

    NASA Astrophysics Data System (ADS)

    Treffman, Lia M.; Morrison, Philip W., Jr.

    1996-04-01

    A process has been developed to hand grind and polish infrared KBr lenses with a wide choice of focal lengths. These lenses are designed for coupling the infrared (IR) beam from a Fourier transform infrared spectrometer into a hot filament chemical vapor deposition reactor. These IR lenses are ground using a glass lens as a mold; the mold lens is a commercial glass lens used for spectacles and is available in a wide range of focal lengths. Rough grinding begins with ♯100 sandpaper and followed by a sequence of fine grinding steps under ethanol: ♯400 SiC powder, ♯600 SiC powder, and emery powder. Polishing is done with a CeO slurry in water followed by a chamois cloth moistened with ethanol. As an indication of the surface finish of the lenses, a flat KBr window that is hand ground and polished in the above manner has a transmission of ≳89% at 8000 cm-1 and ≳91% at 500 cm-1; the commercially polished KBr window has a transmittance of 92% throughout this spectral range.

  6. The Method of Unitary Clothing Transformations in the Theory of Nucleon-Nucleon Scattering

    NASA Astrophysics Data System (ADS)

    Dubovyk, I.; Shebeko, O.

    2010-12-01

    The clothing procedure, put forward in quantum field theory (QFT) by Greenberg and Schweber, is applied for the description of nucleon-nucleon ( N- N) scattering. We consider pseudoscalar ( π and η), vector ( ρ and ω) and scalar ( δ and σ) meson fields interacting with 1/2 spin ( N and {bar{N}}) fermion ones via the Yukawa-type couplings to introduce trial interactions between “bare” particles. The subsequent unitary clothing transformations are found to express the total Hamiltonian through new interaction operators that refer to particles with physical (observable) properties, the so-called clothed particles. In this work, we are focused upon the Hermitian and energy-independent operators for the clothed nucleons, being built up in the second order in the coupling constants. The corresponding analytic expressions in momentum space are compared with the separate meson contributions to the one-boson-exchange potentials in the meson theory of nuclear forces. In order to evaluate the T matrix of the N- N scattering we have used an equivalence theorem that enables us to operate in the clothed particle representation (CPR) instead of the bare particle representation with its large amount of virtual processes. We have derived the Lippmann-Schwinger type equation for the CPR elements of the T-matrix for a given collision energy in the two-nucleon sector of the Hilbert space {mathcal{H}} of hadronic states.

  7. Dark-gray soliton transformations: possibility to study microscopic quantum phenomena by nonlinear optical methods

    NASA Astrophysics Data System (ADS)

    Hernández-Tenorio, C.; Serkin, V. N.; Belyaeva, T. L.; Peña-Moreno, R.; Morales-Lara, L.

    2015-01-01

    The nonlinear Schrödinger equation (NLSE) model with an external harmonic potential is one of the most important in modern science. This model makes it possible to analyze a variety of nonlinear phenomena, in nonlinear optics and laser physics, biophysics and in the theory of Bose-Einstein condensation of atoms. It is shown that the main specific feature of the dynamics of dark GP matter wave solitons in a parabolic trap is the formation of solitons with dynamically changing form-factors producing the periodic variation in the modulation depth (the degree of "blackness") of dark solitons. In general, the period of dark soliton oscillations in trapping potential depends on the specific conditions of the experiment and does not coincide with the oscillation period of a linear quantum-mechanical oscillator. In the case of an immobile pedestal in the trap, the oscillation period of the black soliton considerably increases because of the periodic transformation of the black soliton to the gray one and vice versa. Surprisingly, that if the dark soliton is superimposed on the base pedestal oscillating in the trap and displaced from the trap center, the oscillation period of the dark soliton coincides with the period of oscillations of the linear harmonic oscillator, while the dynamics of the dark soliton is similar to that of a classical particle obeying the Newton mechanics laws.

  8. Optimatization of transient transformation methods to study gene expression in Musa acuminata (AAA group) cultivar Ambon Lumut

    NASA Astrophysics Data System (ADS)

    Prayuni, Kinasih; Dwivany, Fenny M.

    2015-09-01

    Banana is classified as a climateric fruit, whose ripening is regulated by ethylene. Ethylene is synthesized from ACC (1-aminocyclopropane-1-carboxylic acid) by ACC oxidase enzyme which is encoded by ACO gene. Controling an important gene expression in ethylene biosynthesis pathway has became a target to delay the ripening process. Therefore in the previous study we have designed a MaACO-RNAi construct to control MaACO gene expression. In this research, we study the effectiveness of different transient transformation methods to deliver the construct. Direct injection, with or no vaccum infiltration methods were used to deliver MaACO-RNAi construct. All of the methods succesfully deliver the construct into banana fruits based on RT-PCR result.

  9. A Fast and Practical Yeast Transformation Method Mediated by Escherichia coli Based on a Trans-Kingdom Conjugal Transfer System: Just Mix Two Cultures and Wait One Hour

    PubMed Central

    Moriguchi, Kazuki; Yamamoto, Shinji; Ohmine, Yuta; Suzuki, Katsunori

    2016-01-01

    Trans-kingdom conjugation is a phenomenon by which DNA is transferred into a eukaryotic cell by a bacterial conjugal transfer system. Improvement in this method to facilitate the rapid co-cultivation of donor bacterial and recipient eukaryotic cell cultures could make it the simplest transformation method, requiring neither isolation of vector DNA nor preparation of competent recipient cells. To evaluate this potential advantage of trans-kingdom conjugation, we examined this simple transformation method using vector combinations, helper plasmids, and recipient Saccharomyces cerevisiae strains. Mixing donor Escherichia coli and recipient S. cerevisiae overnight cultures (50 μL each) consistently yielded on the order of 101 transformants using the popular experimental strain BY4742 derived from S288c and a shuttle vector for trans-kingdom conjugation. Transformation efficiency increased to the order of 102 using a high receptivity trans-kingdom conjugation strain. In addition, either increasing the amount of donor cells or pretreating the recipient cells with thiols such as dithiothreitol improved the transformation efficiency by one order of magnitude. This simple trans-kingdom conjugation-mediated transformation method could be used as a practical yeast transformation method upon enrichment of available vectors and donor E. coli strains. PMID:26849654

  10. A Benchmark Test Set for Alchemical Free Energy Transformations and Its Use to Quantify Error in Common Free Energy Methods.

    PubMed

    Paliwal, Himanshu; Shirts, Michael R

    2011-12-13

    There is a significant need for improved tools to validate thermophysical quantities computed via molecular simulation. In this paper we present the initial version of a benchmark set of testing methods for calculating free energies of molecular transformation in solution. This set is based on molecular changes common to many molecular design problems, such as insertion and deletion of atomic sites and changing atomic partial charges. We use this benchmark set to compare the statistical efficiency, reliability, and quality of uncertainty estimates for a number of published free energy methods, including thermodynamic integration, free energy perturbation, the Bennett acceptance ratio (BAR) and its multistate equivalent MBAR. We identify MBAR as the consistently best performing method, though other methods are frequently comparable in reliability and accuracy in many cases. We demonstrate that assumptions of Gaussian distributed errors in free energies are usually valid for most methods studied. We demonstrate that bootstrap error estimation is a robust and useful technique for estimating statistical variance for all free energy methods studied. This benchmark set is provided in a number of different file formats with the hope of becoming a useful and general tool for method comparisons.

  11. Unlocking the potential of tropical root crop biotechnology in east Africa by establishing a genetic transformation platform for local farmer-preferred cassava cultivars.

    PubMed

    Nyaboga, Evans; Njiru, Joshua; Nguu, Edward; Gruissem, Wilhelm; Vanderschuren, Herve; Tripathi, Leena

    2013-01-01

    Cassava genetic transformation capacity is still mostly restricted to advanced laboratories in the USA, Europe and China; and its implementation and maintenance in African laboratories has remained scarce. The impact of transgenic technologies for genetic improvement of cassava will depend largely on the transfer of such capabilities to researchers in Africa, where cassava has an important socioeconomic niche. A major constraint to the development of genetic transformation technologies for cassava improvement has been the lack of an efficient and robust transformation and regeneration system. Despite the success achieved in genetic modification of few cassava cultivars, including the model cultivar 60444, transgenic cassava production remains difficult for farmer-preferred cultivars. In this study, a protocol for cultivar 60444 developed at ETH Zurich was successfully implemented and optimized to establish transformation of farmer-preferred cassava cultivars popular in east Africa. The conditions for production and proliferation of friable embryogenic calli (FEC) and Agrobacterium-mediated transformation were optimized for three east African farmer-preferred cultivars (Ebwanatereka, Kibandameno and Serere). Our results demonstrated transformation efficiencies of about 14-22 independent transgenic lines per 100 mg of FEC for farmer-preferred cultivars in comparison to 28 lines per 100 mg of the model cultivar 60444. The presence, integration and expression of the transgenes were confirmed by PCR, Southern blot analysis and histochemical GUS assay. This study reports the establishment of a cassava transformation platform at International Institute of Tropical Agriculture (IITA) hosted by Biosciences eastern and central Africa (BecA) hub in Kenya and provides the basis for transferring important traits such as virus resistance and prolonged shelf-life to farmer-preferred cultivars in east Africa. We anticipate that such platform will also be instrumental to transfer

  12. Unlocking the potential of tropical root crop biotechnology in east Africa by establishing a genetic transformation platform for local farmer-preferred cassava cultivars

    PubMed Central

    Nyaboga, Evans; Njiru, Joshua; Nguu, Edward; Gruissem, Wilhelm; Vanderschuren, Herve; Tripathi, Leena

    2013-01-01

    Cassava genetic transformation capacity is still mostly restricted to advanced laboratories in the USA, Europe and China; and its implementation and maintenance in African laboratories has remained scarce. The impact of transgenic technologies for genetic improvement of cassava will depend largely on the transfer of such capabilities to researchers in Africa, where cassava has an important socioeconomic niche. A major constraint to the development of genetic transformation technologies for cassava improvement has been the lack of an efficient and robust transformation and regeneration system. Despite the success achieved in genetic modification of few cassava cultivars, including the model cultivar 60444, transgenic cassava production remains difficult for farmer-preferred cultivars. In this study, a protocol for cultivar 60444 developed at ETH Zurich was successfully implemented and optimized to establish transformation of farmer-preferred cassava cultivars popular in east Africa. The conditions for production and proliferation of friable embryogenic calli (FEC) and Agrobacterium-mediated transformation were optimized for three east African farmer-preferred cultivars (Ebwanatereka, Kibandameno and Serere). Our results demonstrated transformation efficiencies of about 14–22 independent transgenic lines per 100 mg of FEC for farmer-preferred cultivars in comparison to 28 lines per 100 mg of the model cultivar 60444. The presence, integration and expression of the transgenes were confirmed by PCR, Southern blot analysis and histochemical GUS assay. This study reports the establishment of a cassava transformation platform at International Institute of Tropical Agriculture (IITA) hosted by Biosciences eastern and central Africa (BecA) hub in Kenya and provides the basis for transferring important traits such as virus resistance and prolonged shelf-life to farmer-preferred cultivars in east Africa. We anticipate that such platform will also be instrumental to transfer

  13. A Method for Estimating View Transformations from Image Correspondences Based on the Harmony Search Algorithm.

    PubMed

    Cuevas, Erik; Díaz, Margarita

    2015-01-01

    In this paper, a new method for robustly estimating multiple view relations from point correspondences is presented. The approach combines the popular random sampling consensus (RANSAC) algorithm and the evolutionary method harmony search (HS). With this combination, the proposed method adopts a different sampling strategy than RANSAC to generate putative solutions. Under the new mechanism, at each iteration, new candidate solutions are built taking into account the quality of the models generated by previous candidate solutions, rather than purely random as it is the case of RANSAC. The rules for the generation of candidate solutions (samples) are motivated by the improvisation process that occurs when a musician searches for a better state of harmony. As a result, the proposed approach can substantially reduce the number of iterations still preserving the robust capabilities of RANSAC. The method is generic and its use is illustrated by the estimation of homographies, considering synthetic and real images. Additionally, in order to demonstrate the performance of the proposed approach within a real engineering application, it is employed to solve the problem of position estimation in a humanoid robot. Experimental results validate the efficiency of the proposed method in terms of accuracy, speed, and robustness.

  14. A Method for Estimating View Transformations from Image Correspondences Based on the Harmony Search Algorithm

    PubMed Central

    Cuevas, Erik; Díaz, Margarita

    2015-01-01

    In this paper, a new method for robustly estimating multiple view relations from point correspondences is presented. The approach combines the popular random sampling consensus (RANSAC) algorithm and the evolutionary method harmony search (HS). With this combination, the proposed method adopts a different sampling strategy than RANSAC to generate putative solutions. Under the new mechanism, at each iteration, new candidate solutions are built taking into account the quality of the models generated by previous candidate solutions, rather than purely random as it is the case of RANSAC. The rules for the generation of candidate solutions (samples) are motivated by the improvisation process that occurs when a musician searches for a better state of harmony. As a result, the proposed approach can substantially reduce the number of iterations still preserving the robust capabilities of RANSAC. The method is generic and its use is illustrated by the estimation of homographies, considering synthetic and real images. Additionally, in order to demonstrate the performance of the proposed approach within a real engineering application, it is employed to solve the problem of position estimation in a humanoid robot. Experimental results validate the efficiency of the proposed method in terms of accuracy, speed, and robustness. PMID:26339228

  15. Short-time Fourier transform method for online glass ream inspection

    NASA Astrophysics Data System (ADS)

    Li, Anding; Chen, Youping; Xie, Jingming

    2016-08-01

    In the float glass defect inspection system based on machine vision, the common defects can be detected and classified correctly. However, the online glass ream inspection is still a problem to be solved, and the accuracy cannot fulfill the requirement of glass grading. As the glass ream is a rather slight optical distortion in most cases, it should be amplified so that it can be checked out easily. A Moiré method based on the theory of superposition shadow is proposed, which can magnify the distortion effectively without increasing the system complexity, and a STFT method is presented to analyze the distortion according to the characteristics of Moiré fringe pattern. With this method, the vision interference angle of glass ribbon can be calculated out accurately in real time, which makes it suitable for online glass ream inspection.

  16. A Method for Structure–Activity Analysis of Quorum-Sensing Signaling Peptides from Naturally Transformable Streptococci

    PubMed Central

    2009-01-01

    Many species of streptococci secrete and use a competence-stimulating peptide (CSP) to initiate quorum sensing for induction of genetic competence, bacteriocin production, and other activities. These signaling molecules are small, unmodified peptides that induce powerful strain-specific activity at nano-molar concentrations. This feature has provided an excellent opportunity to explore their structure–function relationships. However, CSP variants have also been identified in many species, and each specifically activates its cognate receptor. How such minor changes dramatically affect the specificity of these peptides remains unclear. Structure–activity analysis of these peptides may provide clues for understanding the specificity of signaling peptide–receptor interactions. Here, we use the Streptococcus mutans CSP as an example to describe methods of analyzing its structure–activity relationship. The methods described here may provide a platform for studying quorum-sensing signaling peptides of other naturally transformable streptococci. PMID:19517207

  17. [A study of coordinates transform iterative fitting method to extract bio-impedance model parameters bio-impedance model parameters].

    PubMed

    Zhou, Liming; Yang, Yuxing; Yuan, Shiying

    2006-02-01

    A new algorithm, the coordinates transform iterative optimizing method based on the least square curve fitting model, is presented. This arithmetic is used for extracting the bio-impedance model parameters. It is superior to other methods, for example, its speed of the convergence is quicker, and its calculating precision is higher. The objective to extract the model parameters, such as Ri, Re, Cm and alpha, has been realized rapidly and accurately. With the aim at lowering the power consumption, decreasing the price and improving the price-to-performance ratio, a practical bio-impedance measure system with double CPUs has been built. It can be drawn from the preliminary results that the intracellular resistance Ri increased largely with an increase in working load during sitting, which reflects the ischemic change of lower limbs.

  18. Robust Scale Transformation Methods in IRT True Score Equating under Common-Item Nonequivalent Groups Design

    ERIC Educational Resources Information Center

    He, Yong

    2013-01-01

    Common test items play an important role in equating multiple test forms under the common-item nonequivalent groups design. Inconsistent item parameter estimates among common items can lead to large bias in equated scores for IRT true score equating. Current methods extensively focus on detection and elimination of outlying common items, which…

  19. Rapid deletion plasmid construction methods for protoplast and Agrobacterium based fungal transformation systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Increasing availability of genomic data and sophistication of analytical methodology in fungi has elevated the need for functional genomics tools in these organisms. Gene deletion is a critical tool for functional analysis. The targeted deletion of genes requires both a suitable method for the trans...

  20. Harnessing the power of big data: infusing the scientific method with machine learning to transform ecology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Most efforts to harness the power of big data for ecology and environmental sciences focus on data and metadata sharing, standardization, and accuracy. However, many scientists have not accepted the data deluge as an integral part of their research because the current scientific method is not scalab...

  1. Wearable Sensor-Based Human Activity Recognition Method with Multi-Features Extracted from Hilbert-Huang Transform

    PubMed Central

    Xu, Huile; Liu, Jinyi; Hu, Haibo; Zhang, Yi

    2016-01-01

    Wearable sensors-based human activity recognition introduces many useful applications and services in health care, rehabilitation training, elderly monitoring and many other areas of human interaction. Existing works in this field mainly focus on recognizing activities by using traditional features extracted from Fourier transform (FT) or wavelet transform (WT). However, these signal processing approaches are suitable for a linear signal but not for a nonlinear signal. In this paper, we investigate the characteristics of the Hilbert-Huang transform (HHT) for dealing with activity data with properties such as nonlinearity and non-stationarity. A multi-features extraction method based on HHT is then proposed to improve the effect of activity recognition. The extracted multi-features include instantaneous amplitude (IA) and instantaneous frequency (IF) by means of empirical mode decomposition (EMD), as well as instantaneous energy density (IE) and marginal spectrum (MS) derived from Hilbert spectral analysis. Experimental studies are performed to verify the proposed approach by using the PAMAP2 dataset from the University of California, Irvine for wearable sensors-based activity recognition. Moreover, the effect of combining multi-features vs. a single-feature are investigated and discussed in the scenario of a dependent subject. The experimental results show that multi-features combination can further improve the performance measures. Finally, we test the effect of multi-features combination in the scenario of an independent subject. Our experimental results show that we achieve four performance indexes: recall, precision, F-measure, and accuracy to 0.9337, 0.9417, 0.9353, and 0.9377 respectively, which are all better than the achievements of related works. PMID:27918414

  2. Wearable Sensor-Based Human Activity Recognition Method with Multi-Features Extracted from Hilbert-Huang Transform.

    PubMed

    Xu, Huile; Liu, Jinyi; Hu, Haibo; Zhang, Yi

    2016-12-02

    Wearable sensors-based human activity recognition introduces many useful applications and services in health care, rehabilitation training, elderly monitoring and many other areas of human interaction. Existing works in this field mainly focus on recognizing activities by using traditional features extracted from Fourier transform (FT) or wavelet transform (WT). However, these signal processing approaches are suitable for a linear signal but not for a nonlinear signal. In this paper, we investigate the characteristics of the Hilbert-Huang transform (HHT) for dealing with activity data with properties such as nonlinearity and non-stationarity. A multi-features extraction method based on HHT is then proposed to improve the effect of activity recognition. The extracted multi-features include instantaneous amplitude (IA) and instantaneous frequency (IF) by means of empirical mode decomposition (EMD), as well as instantaneous energy density (IE) and marginal spectrum (MS) derived from Hilbert spectral analysis. Experimental studies are performed to verify the proposed approach by using the PAMAP2 dataset from the University of California, Irvine for wearable sensors-based activity recognition. Moreover, the effect of combining multi-features vs. a single-feature are investigated and discussed in the scenario of a dependent subject. The experimental results show that multi-features combination can further improve the performance measures. Finally, we test the effect of multi-features combination in the scenario of an independent subject. Our experimental results show that we achieve four performance indexes: recall, precision, F-measure, and accuracy to 0.9337, 0.9417, 0.9353, and 0.9377 respectively, which are all better than the achievements of related works.

  3. Cyclic voltammetry as a sensitive method for in situ probing of chemical transformations in quantum dots.

    PubMed

    Osipovich, Nikolai P; Poznyak, Sergei K; Lesnyak, Vladimir; Gaponik, Nikolai

    2016-04-21

    The application of electrochemical methods for the characterization of colloidal quantum dots (QDs) attracts considerable attention as these methods may allow for monitoring of some crucial parameters, such as energetic levels of conduction and valence bands as well as surface traps and ligands under real conditions of colloidal solution. In the present work we extend the applications of cyclic voltammetry (CV) to in situ monitoring of degradation processes of water-soluble CdTe QDs. This degradation occurs under lowering of pH to the values around 5, i.e. under conditions relevant to bioimaging applications of these QDs, and is accompanied by pronounced changes of their photoluminescence. Observed correlations between characteristic features of CV diagrams and the fluorescence spectra allowed us to propose mechanisms responsible for evolution of the photoluminescence properties as well as degradation pathway of CdTe QDs at low pH.

  4. An implicit fast Fourier transform method for integration of the time dependent Schrodinger equation

    SciTech Connect

    Riley, M.E.; Ritchie, A.B.

    1997-12-31

    One finds that the conventional exponentiated split operator procedure is subject to difficulties when solving the time-dependent Schrodinger equation for Coulombic systems. By rearranging the kinetic and potential energy terms in the temporal propagator of the finite difference equations, one can find a propagation algorithm for three dimensions that looks much like the Crank-Nicholson and alternating direction implicit methods for one- and two-space-dimensional partial differential equations. The authors report investigations of this novel implicit split operator procedure. The results look promising for a purely numerical approach to certain electron quantum mechanical problems. A charge exchange calculation is presented as an example of the power of the method.

  5. [Research on the application of response variable transform method in high scattering medium with near-infrared spectroscopy].

    PubMed

    Peng, Dan; Xu, Ke-Xin

    2010-02-01

    In the present paper, a new method named response variable transform (RVT) is proposed to reduce the effect of scattering in the NIR spectroscopy measurement. In the RVT method, the rates of the change in light intensity at different spatial positions with the concentration of the investigated composition are firstly sampled. Then, based on the sampled changing rates of light intensity, the absorption coefficients, which are used as new response variable, are calculated using the inverse adding-double (IAD) algorithm. Finally, through the partial least squares (PLS) regression, the calibration models for the investigated compositions are established using these new response variables. To validate the feasibility and effectiveness of the proposed method, the RVT method is applied to reduce the effect of scattering during the process of composition concentration measurement in milk. First, the relationships between the absorption coefficient and the concentrations of major compositions including fat and protein are analyzed. Then, with Monte Carlo simulations for different concentration levels of fat and protein, the influence of scattering on the relationship between the absorbency and the concentration is investigated and verified. Finally, for comparison, both the RVT method and the traditional preprocessing techniques are used to reduce the effect of scattering during the process of prediction model establishment. Experimental results showed that the prediction precision for fat and protein with the RVT method is increased by up to 33.9% and 46.5% than that with multiplicative signal correction (MSC) algorithm, and the prediction precision for fat and protein with the RVT method is increased by up to 55.9% and 33.7% than that with standard normal variate (SNV) algorithm, which also indicates that the RVT method is an effective method for eliminating the interference of scattering in the NIR spectroscopy measurement.

  6. A quantitative method to detect explosives and other selected semivolatiles in soil samples by Fourier transform infrared spectroscopy

    SciTech Connect

    Clapper-Gowdy, M.; Demirgian, J.; Lang, K.; Robaittaille, G.

    1992-09-01

    The current methods for hazardous waste site characterization are time consuming, cumbersome, and expensive. Typically, characterization requires a preliminary site assessment and subsequent sampling of potentially contaminated soils and waters. The samples are sent to laboratories for analysis using EPA-certified methods. It is often necessary to repeat the entire sampling-analysis cycle to characterize a site completely and accurately. For these reasons, new methods of site assessment and characterization are continually being researched. TWs paper describes a Fourier transform infrared (FTIR) spectroscopy method that rapidly screens soil samples from potentially hazardous waste sites. Analysis of a soil sample by FTIR takes approximately 10 minutes. The method has been developed to identify and quantify explosives in the field and is directly applicable to selected volatile organics, semivolatile organics, and pesticides. The soil samples are desorbed in a CDS 122 thermal desorption unit under vacuum into a variable pathlength, long-path cell heated to 180{degrees}C. The spectral data, 128 co-added scans at I cm{sup {minus}l} resolution, are collected and stored using a Nicolet 60SX FTIR spectrometer. Classical least squares (CLS) analysis has been used to obtain quantitative results.

  7. A quantitative method to detect explosives and other selected semivolatiles in soil samples by Fourier transform infrared spectroscopy

    SciTech Connect

    Clapper-Gowdy, M.; Demirgian, J. ); Lang, K.; Robaittaille, G. )

    1992-01-01

    The current methods for hazardous waste site characterization are time consuming, cumbersome, and expensive. Typically, characterization requires a preliminary site assessment and subsequent sampling of potentially contaminated soils and waters. The samples are sent to laboratories for analysis using EPA-certified methods. It is often necessary to repeat the entire sampling-analysis cycle to characterize a site completely and accurately. For these reasons, new methods of site assessment and characterization are continually being researched. TWs paper describes a Fourier transform infrared (FTIR) spectroscopy method that rapidly screens soil samples from potentially hazardous waste sites. Analysis of a soil sample by FTIR takes approximately 10 minutes. The method has been developed to identify and quantify explosives in the field and is directly applicable to selected volatile organics, semivolatile organics, and pesticides. The soil samples are desorbed in a CDS 122 thermal desorption unit under vacuum into a variable pathlength, long-path cell heated to 180{degrees}C. The spectral data, 128 co-added scans at I cm{sup {minus}l} resolution, are collected and stored using a Nicolet 60SX FTIR spectrometer. Classical least squares (CLS) analysis has been used to obtain quantitative results.

  8. A comparative study between Empirical Wavelet Transforms and Empirical Mode Decomposition Methods: Application to bearing defect diagnosis

    NASA Astrophysics Data System (ADS)

    Kedadouche, M.; Thomas, M.; Tahan, A.

    2016-12-01

    The Ensemble Empirical Mode Decomposition (EEMD) is a noise assisted method that may sometimes provide a significant improvement on Empirical Mode Decomposition (EMD). However, the amplitude and number of added noise need to be selected when applying the EEMD method. Furthermore, the computation time which depends on the number of ensemble trails is very high compared to the EMD. In this paper, a new way for choosing the appropriate added noise is presented. Conversely, a recently-developed method called the Empirical Wavelet Transform (EWT) is investigated. A comparative study between the EMD and EWT methods is conducted. The results show that the EWT is better than the EEMD and EMD on mode estimates and computation time is significantly reduced. An experimental study on bearing diagnosis is conducted. The EWT is applied to experimental data coming from damaged bearings. In the paper, an index selection is introduced that allows for the automatic selection of the Intrinsic Mode Functions (IMF) that should be used to perform the envelope spectrum. It is shown that choosing all the IMF selected by the index is more efficient than only choosing the best one. The envelope of the sum of the selected IMF clearly reveals the bearing frequencies and its harmonics which are excited by the defect. This approach seems to be an effective and efficient method for processing bearing fault signals.

  9. Autocorrelation Analysis Combined with a Wavelet Transform Method to Detect and Remove Cosmic Rays in a Single Raman Spectrum.

    PubMed

    Maury, Augusto; Revilla, Reynier I

    2015-08-01

    Cosmic rays (CRs) occasionally affect charge-coupled device (CCD) detectors, introducing large spikes with very narrow bandwidth in the spectrum. These CR features can distort the chemical information expressed by the spectra. Consequently, we propose here an algorithm to identify and remove significant spikes in a single Raman spectrum. An autocorrelation analysis is first carried out to accentuate the CRs feature as outliers. Subsequently, with an adequate selection of the threshold, a discrete wavelet transform filter is used to identify CR spikes. Identified data points are then replaced by interpolated values using the weighted-average interpolation technique. This approach only modifies the data in a close vicinity of the CRs. Additionally, robust wavelet transform parameters are proposed (a desirable property for automation) after optimizing them with the application of the method in a great number of spectra. However, this algorithm, as well as all the single-spectrum analysis procedures, is limited to the cases in which CRs have much narrower bandwidth than the Raman bands. This might not be the case when low-resolution Raman instruments are used.

  10. Methods for suspension culture, protoplast extraction, and transformation of high-biomass yielding perennial grass Arundo donax.

    PubMed

    Pigna, Gaia; Dhillon, Taniya; Dlugosz, Elizabeth M; Yuan, Joshua S; Gorman, Connor; Morandini, Piero; Lenaghan, Scott C; Stewart, C Neal

    2016-12-01

    Arundo donax L. is a promising biofuel feedstock in the Mediterranean region. Despite considerable interest in its genetic improvement, Arundo tissue culture and transformation remains arduous. The authors developed methodologies for cell- and tissue culture and genetic engineering in Arundo. A media screen was conducted, and a suspension culture was established using callus induced from stem axillary bud explants. DBAP medium, containing 9 µM 2,4-D and 4.4 µM BAP, was found to be the most effective medium among those tested for inducing cell suspension cultures, which resulted in a five-fold increase in tissue mass over 14 days. In contrast, CIM medium containing 13 µM 2,4-D, resulted in just a 1.4-fold increase in mass over the same period. Optimized suspension cultures were superior to previously-described solidified medium-based callus culture methods for tissue mass increase. Suspension cultures proved to be very effective for subsequent protoplast isolation. Protoplast electroporation resulted in a 3.3 ± 1.5% transformation efficiency. A dual fluorescent reporter gene vector enabled the direct comparison of the CAMV 35S promoter with the switchgrass ubi2 promoter in single cells of Arundo. The switchgrass ubi2 promoter resulted in noticeably higher reporter gene expression compared with that conferred by the 35S promoter in Arundo.

  11. Near UV-near IR Fourier transform spectrometer using the beam-folding position-tracking method based on retroreflectors.

    PubMed

    Wang, Xuzhu; Chan, Robert K Y; Cheng, Amelia S K

    2008-12-01

    A near UV-near IR Fourier transform spectrometer based on a beam-folding position-tracking method realized by using retroreflectors is reported. The use of retroreflectors maintains all beams in the beam-fold arrangement in parallel with the incident beams. The beam-folding interferometer used for position tracking is arranged to have optical path symmetry with the measurement interferometer in the zero path difference position of the measurement interferometer, and the vertex of the movable retroreflector in the measurement interferometer is arranged very close to the midpoint of the vertices of two movable retroreflectors in the position-tracking interferometer. These measures keep the equivalent optical axis of the position-tracking interferometer well in line with that of the measurement interferometer even with translational misalignments. Therefore, the change in the optical path difference of the position-tracking interferometer is always synchronous to that of the measurement interferometer during the scanning process. That is, the position-tracking error can be suppressed to very small values during a scan. We have demonstrated a UV-near IR Fourier transform spectrometer with a standard quality ball-bearing translation stage achieving a resolution close to the theoretical resolution of approximately 0.28 cm(-1) at the He-Ne laser wavelength when the scan distance reaches the travel distance of over 2 cm. This was achieved without the need for elaborate optics, sophisticated detecting electronics, and high-precision servomotion control.

  12. Wide-stripe noise removal method of hyperspectral image based on fusion of wavelet transform and local interpolation

    NASA Astrophysics Data System (ADS)

    Huang, Shiqi; Liu, Zhigang; Wang, Yiting; Wang, Rongrong

    2017-03-01

    The principle of hyperspectral imaging leads to a variety of stripe noise in hyperspectral images, especially the wide-stripe noise, which brings great obstacles to the interpretation and application of hyperspectral images. Aiming at the wide-stripe noise of hyperspectral images of two-level production data, considering from the effect of filtering noise and the ability of protecting detail, this paper proposed a fused wide-stripe removal method based on the wavelet transform and local interpolation (WTLI), called the WTLI algorithm. On one hand, it uses the wavelet transform to remove the stripe noise as much as possible; on the other hand, it uses the local interpolation to protect more geometric and detailed information, so as to achieve the purpose of removing noise and protecting the useful information. A series of comparative experiments were carried out with hyperspectral image data. Not only have good experimental results been obtained, but also this shows that the WTLI algorithm has better stability and universality.

  13. A New Method for the Identification of Surfactant-Producing or Transforming Bacteria in the Sea Surface Microlayer

    NASA Astrophysics Data System (ADS)

    Vella, K.; Kurata, N.; Soloviev, A.; Matt, S.; Shivji, M.; Tartar, A.

    2012-12-01

    The sea surface microlayer represents the interface between the atmosphere and the ocean and it can serve as a collection point for organic materials, such as surfactants. In this work, we present an improved method for the identification of the microbial community structure present in both the sea surface microlayer and the corresponding water column, in relation to surfactant production/transformation. Bacterial samples were collected from the sea surface microlayer with an improved sampling method, which minimized sample contamination from the research platform and the subsurface water. This novel sampling approach was developed by using polycarbonate membrane filters attached to fly fishing line, which provided finer control of the filter touching only the sea surface. Sampling was conducted at open ocean (approximately five miles offshore) and coastal water sites (approximately one mile from coastline) in the Straits of Florida to compare surfactant-producing bacterial populations present in these two regions. The microlayer sampling was planned to coincide with synthetic aperture radar (SAR) satellite overpasses (COSMO SkyMed). By establishing bacterial groups that influence the presence of surfactants, remote sensing techniques which involve monitoring the surface of the ocean will be enhanced. This is because surfactants are capable of suppressing short gravity-capillary ocean surface waves, which affect radar back-scatter. SAR remote sensing thus may provide additional information on the state of the upper ocean ecosystem. Microbial composition was determined using DNA analysis of 16S rRNA genes. Preliminary analysis of bay samples suggested the presence of bacteria related to a strain of uncultured gammaproteobacteria, previously isolated from coastal sediment along a hydrocarbon contamination gradient. Many genera of the gammaproteobacteria are known to be involved in hydrocarbon degradation. We are further exploring and quantifying the presence of

  14. Stability analysis of phonon transport equations derived via the Chapman-Enskog method and transformation of variables.

    PubMed

    Banach, Zbigniew; Larecki, Wieslaw; Zajaczkowski, Wojciech

    2009-10-01

    Under the assumption of Callaway's model of the Boltzmann-Peierls equation, the Chapman-Enskog method for a phonon gas forms the basis to derive various hydrodynamic equations for the energy density and the drift velocity of interest when normal processes dominate over resistive ones. The first three levels of the expansion (i.e., the zeroth-, first-, and second-order approximations) are satisfactory in that they are entropy consistent and ensure linear stability of the rest state. However, the entropy density contains a weakly nonlocal term, the entropy production is a degenerate function of variables, and the next order in the Chapman-Enskog expansion gives the equations with linearly unstable rest solutions. In the context of Burnett and super-Burnett equations, a similar type of problem was recognized by several authors who proposed different ways to deal with it. Here we report on yet another possible device for obtaining more satisfactory equations. Namely, inspired by the fact that there exists no unique way to truncate the Chapman-Enskog expansion, we combine the Chapman-Enskog procedure with the method of variable transformation and subsequently find a class of epsilon -dependent transformations through which it is possible to derive the second-order equations possessing a local entropy density and nondegenerate expression for the entropy production. Regardless of this result, we also show that although the method cannot be used to construct linearly stable third-order equations, it can be used to make the originally stable first-order equations asymptotically stable.

  15. Optimization of in vitro regeneration and Agrobacterium tumefaciens-mediated transformation with heat-resistant cDNA in Brassica oleracea subsp. italica cv. Green Marvel.

    PubMed

    Ravanfar, Seyed Ali; Aziz, Maheran Abdul; Saud, Halimi Mohd; Abdullah, Janna Ong

    2015-11-01

    An efficient system for shoot regeneration and Agrobacterium tumefaciens-mediated transformation of Brassica oleracea cv. Green Marvel cultivar is described. This study focuses on developing shoot regeneration from hypocotyl explants of broccoli cv. Green Marvel using thidiazuron (TDZ), zeatin, and kinetin, the optimization of factors affecting Agrobacterium-mediated transformation of the hypocotyl explants with heat-resistant cDNA, followed by the confirmation of transgenicity of the regenerants. High shoot regeneration was observed in 0.05-0.1 mg dm(-3) TDZ. TDZ at 0.1 mg dm(-3) produced among the highest percentage of shoot regeneration (96.67 %) and mean number of shoot formation (6.17). The highest percentage (13.33 %) and mean number (0.17) of putative transformant production were on hypocotyl explants subjected to preculture on shoot regeneration medium (SRM) with 200 µM acetosyringone. On optimization of bacterial density and inoculation time, the highest percentage and mean number of putative transformant production were on hypocotyl explants inoculated with a bacterial dilution of 1:5 for 30 min. Polymerase chain reaction (PCR) assay indicated a transformation efficiency of 8.33 %. The luciferase assay showed stable integration of the Arabidopsis thaliana HSP101 (AtHSP101) cDNA in the transgenic broccoli regenerants. Three out of five transgenic lines confirmed through PCR showed positive hybridization bands of the AtHSP101 cDNA through Southern blot analysis. The presence of AtHSP101 transcripts in the three transgenic broccoli lines indicated by reverse transcription-PCR (RT-PCR) confirmed the expression of the gene. In conclusion, an improved regeneration system has been established from hypocotyl explants of broccoli followed by successful transformation with AtHSP101 for resistance to high temperature.

  16. CULTURALLY COMPETENT SERVICES WITHIN A STATEWIDE BEHAVIORAL HEALTHCARE TRANSFORMATION: A MIXED-METHOD ASSESSMENT

    PubMed Central

    Semansky, Rafael M.; Goodkind, Jessica; Sommerfeld, David H.; Willging, Cathleen E.

    2014-01-01

    In 2005, New Mexico created a single health plan to administer all publicly-funded behavioral health services. Our mixed-method study combined surveys, document review, and ethnography to examine this reform’s influence on culturally competent services (CCS). Participants were executives, providers, and support staff of behavioral healthcare agencies. Key variables included language access services and organizational supports, i.e., training, self-assessments of CCS, and maintenance of client-level data. Survey and document review suggested minimal effects on statewide capacity for CCS during the first three years of the reform. Ethnographic research helped explain these findings: (1) state government, the managed behavioral health plan and agencies failed to champion CCS; and (2) increased administrative requirements minimized time and financial resources for CCS. There was also insufficient appreciation among providers for CCS. Although agencies made progress in addressing language assistance services, availability and quality remained limited. PMID:25937679

  17. Isochoric Burn, an Internally Consistent Method for the Reactant to Product Transformation in Reactive Flow

    SciTech Connect

    Reaugh, J E; Lee, E L

    2002-07-01

    Mixture rules for partially reacted explosives differ amongst various models. For instance, JWL++ uses a partial pressure addition to compute an average zonal pressure, Ignition and Growth requires pressure equilibration and thermal equilibration of temperature dependent JWL EOSs, CHEETAH In Line RF also assumes temperature and pressure equilibration. It has been suggested in the past that a more realistic equilibration scheme should comprise isentropic pressure equilibration of the separate reacted and unreacted phases. This turns out not to be a proper path for equilibration. Rather, we find that the only internally consistent method is the evaluation of the equilibrium pressure that satisfies the particular conditions of reactant and product resulting from deflagration in a fixed volume.

  18. Rapid method for determining malondialdehyde as secondary oxidation product in palm olein system by Fourier transform infrared spectroscopy.

    PubMed

    Mirghani, M E S; Che Man, Y B; Jinap, S; Baharin, B S; Bakar, J

    2002-01-01

    A simple and rapid Fourier transform infrared (FTIR) spectroscopic method has been developed for the quantitative determination of malondialdehyde as secondary oxidation product in a palm olein system. The FTIR method was based on a sodium chloride transmission cell and utilised a partial least square statistical approach to derive a calibration model. The frequency region combinations that gave good calibration were 2900-2800, and 1800-1600 cm-1. The precision and accuracy, in the range 0-60 mumol malondialdehyde/kg oil, were comparable to those of the modified distillation method with a coefficient of determination (r2) of 0.9891 and standard error of calibration of 1.49. The calibration was cross-validated and produced an r2 of 0.9786 and standard error of prediction of 2.136. The results showed that the FTIR method is versatile, efficient and accurate, and suitable for routine quality control analysis with the result obtainable in about 2 min from a sample of less than 2 mL.

  19. Continuous wavelet transform and Euler deconvolution method and their application to magnetic field data of Jharia coalfield, India

    NASA Astrophysics Data System (ADS)

    Singh, Arvind; Singh, Upendra Kumar

    2017-02-01

    This paper deals with the application of continuous wavelet transform (CWT) and Euler deconvolution methods to estimate the source depth using magnetic anomalies. These methods are utilized mainly to focus on the fundamental issue of mapping the major coal seam and locating tectonic lineaments. The main aim of the study is to locate and characterize the source of the magnetic field by transferring the data into an auxiliary space by CWT. The method has been tested on several synthetic source anomalies and finally applied to magnetic field data from Jharia coalfield, India. Using magnetic field data, the mean depth of causative sources points out the different lithospheric depth over the study region. Also, it is inferred that there are two faults, namely the northern boundary fault and the southern boundary fault, which have an orientation in the northeastern and southeastern direction respectively. Moreover, the central part of the region is more faulted and folded than the other parts and has sediment thickness of about 2.4 km. The methods give mean depth of the causative sources without any a priori information, which can be used as an initial model in any inversion algorithm.

  20. A new method for determining the relative crystallinity of chickpea starch by Fourier-transform infrared spectroscopy.

    PubMed

    Sun, Yongkang; Wu, Zhongwei; Hu, Bing; Wang, Wei; Ye, Hong; Sun, Yi; Wang, Xiaoqing; Zeng, Xiaoxiong

    2014-08-08

    A new method for determining the relative crystallinity (RC) of chickpea starch was developed by using Fourier-transform infrared (FT-IR) spectroscopy, based on hypotheses as described as follows: there is a Gaussian holocrystalline-peak (HCP) in the 800-1,300 cm(-1) region of FT-IR spectrum of starch which is divided into amorphous region and crystalline region; the crystalline region of HCP is the overlap of the HCP and the FT-IR spectrum of starch; the RC of starch is the ratio of the area of crystalline region to the area of HCP. It was found that there was no significant difference between the RC determined by FT-IR method and that determined by X-ray diffraction (XRD) method. The intra-class correlation coefficient was 0.998 (p=0.000, n=9) and the 95% confidence interval was 0.992-1.000 for the RC determined by XRD and FT-IR. Furthermore, the developed method showed good repeatability (coefficient of variation (CV), 1.1-2.9%) and good intermediate precision (CV, 2.8%).